https://referensmetodik.folkhalsomyndigheten.se/index.php?action=history&feed=atom&title=Bilagor_till_VRE-diagnostiken Bilagor till VRE-diagnostiken - Versionshistorik 2026-05-14T04:29:19Z Versionshistorik för denna sida på wikin MediaWiki 1.35.8 https://referensmetodik.folkhalsomyndigheten.se/index.php?title=Bilagor_till_VRE-diagnostiken&diff=12163&oldid=prev Magnus Thore: /* PCR-metod för påvisande av van-gener */ 2012-07-19T10:33:35Z <p><span dir="auto"><span class="autocomment">PCR-metod för påvisande av van-gener</span></span></p> <table class="diff diff-contentalign-left diff-editfont-monospace" data-mw="interface"> <col class="diff-marker" /> <col class="diff-content" /> <col class="diff-marker" /> <col class="diff-content" /> <tr class="diff-title" lang="sv"> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Äldre version</td> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Versionen från 19 juli 2012 kl. 10.33</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l105" >Rad 105:</td> <td colspan="2" class="diff-lineno">Rad 105:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>[[Kategori:Laboratoriediagnostik]]</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>[[Kategori:Laboratoriediagnostik]]</div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">[[Kategori:Smittskyddslagens sjukdomar]]</ins></div></td></tr> </table> Magnus Thore https://referensmetodik.folkhalsomyndigheten.se/index.php?title=Bilagor_till_VRE-diagnostiken&diff=12162&oldid=prev Magnus Thore: /* 
Anrikningsmedium för VRE-screening */ 2012-07-19T10:32:50Z <p><span dir="auto"><span class="autocomment">
Anrikningsmedium för VRE-screening</span></span></p> <table class="diff diff-contentalign-left diff-editfont-monospace" data-mw="interface"> <col class="diff-marker" /> <col class="diff-content" /> <col class="diff-marker" /> <col class="diff-content" /> <tr class="diff-title" lang="sv"> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Äldre version</td> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Versionen från 19 juli 2012 kl. 10.32</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l50" >Rad 50:</td> <td colspan="2" class="diff-lineno">Rad 50:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Todd-Hewittbuljong tillreds enligt tillverkarens anvisningar. Tillsätt vankomycin 4 mg/L och aztreonam 60 mg/L.</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Todd-Hewittbuljong tillreds enligt tillverkarens anvisningar. Tillsätt vankomycin 4 mg/L och aztreonam 60 mg/L.</div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;"></ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">----------------</ins></div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>===Bilaga 3===</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>===Bilaga 3===</div></td></tr> </table> Magnus Thore https://referensmetodik.folkhalsomyndigheten.se/index.php?title=Bilagor_till_VRE-diagnostiken&diff=12161&oldid=prev Magnus Thore: /* Diagnostiska minimikriterier
 för artdiagnostik 
av enterokocker (referensmetodik) */ 2012-07-19T10:32:28Z <p><span dir="auto"><span class="autocomment">Diagnostiska minimikriterier
 för artdiagnostik 
av enterokocker (referensmetodik)</span></span></p> <table class="diff diff-contentalign-left diff-editfont-monospace" data-mw="interface"> <col class="diff-marker" /> <col class="diff-content" /> <col class="diff-marker" /> <col class="diff-content" /> <tr class="diff-title" lang="sv"> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Äldre version</td> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Versionen från 19 juli 2012 kl. 10.32</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l14" >Rad 14:</td> <td colspan="2" class="diff-lineno">Rad 14:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>[[Fil:Tabell 4 enterokocker 0001.jpg|thumb|center|700px||]]</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>[[Fil:Tabell 4 enterokocker 0001.jpg|thumb|center|700px||]]</div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;"></ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">------------</ins></div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>===Bilaga 2===</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>===Bilaga 2===</div></td></tr> </table> Magnus Thore https://referensmetodik.folkhalsomyndigheten.se/index.php?title=Bilagor_till_VRE-diagnostiken&diff=12160&oldid=prev Magnus Thore: /* Diagnostiska minimikriterier
 för artdiagnostik 
av enterokocker (referensmetodik) */ 2012-07-19T10:31:24Z <p><span dir="auto"><span class="autocomment">Diagnostiska minimikriterier
 för artdiagnostik 
av enterokocker (referensmetodik)</span></span></p> <table class="diff diff-contentalign-left diff-editfont-monospace" data-mw="interface"> <col class="diff-marker" /> <col class="diff-content" /> <col class="diff-marker" /> <col class="diff-content" /> <tr class="diff-title" lang="sv"> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Äldre version</td> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Versionen från 19 juli 2012 kl. 10.31</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l9" >Rad 9:</td> <td colspan="2" class="diff-lineno">Rad 9:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>====Diagnostiska minimikriterier
 för artdiagnostik 
av enterokocker (referensmetodik)====</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>====Diagnostiska minimikriterier
 för artdiagnostik 
av enterokocker (referensmetodik)====</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>*&#039;&#039;Enterococcus&#039;&#039; spp: Minimikriterier:
 Typisk kolonimorfologi<del class="diffchange diffchange-inline">, grampositiv
, katalasnegativ, galla-eskulin-positiv, alternativt positiv i agglutinationstest för grupp D, PYR-positiv
. Andra egenskaper: Gråvita </del>kolonier på blodagar, kan ha ingen hemolys, alfa- eller betahemolys, oftast gula kolonier på CLED-agar. <del class="diffchange diffchange-inline">Växer </del>i buljong med 6,5 % NaCl <del class="diffchange diffchange-inline">
</del>(&#039;&#039;&#039;Tabell 4&#039;&#039;&#039;)</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>*&#039;&#039;Enterococcus&#039;&#039; spp: Minimikriterier:
 Typisk kolonimorfologi <ins class="diffchange diffchange-inline">(gråvita </ins>kolonier på blodagar, kan ha ingen hemolys, alfa- eller betahemolys, oftast gula kolonier på CLED-agar<ins class="diffchange diffchange-inline">), katalasnegativa grampositiva kocker i par eller korta kedjor, PYR- och eskulin-positiva</ins>. <ins class="diffchange diffchange-inline">Ytterligare egenskaper för enterokocker är förmåga att växa </ins>i buljong med 6,5 % NaCl<ins class="diffchange diffchange-inline">, positiv i agglutinationstest för Lancefields grupp D. </ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins class="diffchange diffchange-inline">För diffentiering av enterokocker se
</ins>(&#039;&#039;&#039;Tabell 4&#039;&#039;&#039;)</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>[[Fil:Tabell 4 enterokocker 0001.jpg|thumb|center|700px||]]</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>[[Fil:Tabell 4 enterokocker 0001.jpg|thumb|center|700px||]]</div></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del style="font-weight: bold; text-decoration: none;"></del></div></td><td colspan="2"> </td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>===Bilaga 2===</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>===Bilaga 2===</div></td></tr> </table> Magnus Thore https://referensmetodik.folkhalsomyndigheten.se/index.php?title=Bilagor_till_VRE-diagnostiken&diff=8203&oldid=prev Magnus Thore: /* PCR-metod för påvisande av van-gener */ 2012-02-26T09:43:35Z <p><span dir="auto"><span class="autocomment">PCR-metod för påvisande av van-gener</span></span></p> <table class="diff diff-contentalign-left diff-editfont-monospace" data-mw="interface"> <col class="diff-marker" /> <col class="diff-content" /> <col class="diff-marker" /> <col class="diff-content" /> <tr class="diff-title" lang="sv"> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Äldre version</td> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Versionen från 26 februari 2012 kl. 09.43</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l99" >Rad 99:</td> <td colspan="2" class="diff-lineno">Rad 99:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Avläsning: Metoden är en cybergreenmetod och läses av i våglängd 530 nm. Positiva prov ska uppvisa en amplifieringskurva samt att produkten ska ha en smälttopp runt 84–85 °C.</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Avläsning: Metoden är en cybergreenmetod och läses av i våglängd 530 nm. Positiva prov ska uppvisa en amplifieringskurva samt att produkten ska ha en smälttopp runt 84–85 °C.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>[[Kategori:<del class="diffchange diffchange-inline">Laboratoriemetodik</del>]]</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>[[Kategori:<ins class="diffchange diffchange-inline">Laboratoriediagnostik</ins>]]</div></td></tr> </table> Magnus Thore https://referensmetodik.folkhalsomyndigheten.se/index.php?title=Bilagor_till_VRE-diagnostiken&diff=8202&oldid=prev Magnus Thore den 26 februari 2012 kl. 09.42 2012-02-26T09:42:33Z <p></p> <table class="diff diff-contentalign-left diff-editfont-monospace" data-mw="interface"> <col class="diff-marker" /> <col class="diff-content" /> <col class="diff-marker" /> <col class="diff-content" /> <tr class="diff-title" lang="sv"> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Äldre version</td> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Versionen från 26 februari 2012 kl. 09.42</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l58" >Rad 58:</td> <td colspan="2" class="diff-lineno">Rad 58:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>[[Fil:VanAochvanB_LightCycler.jpg|thumb|center|600px||]]</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>[[Fil:VanAochvanB_LightCycler.jpg|thumb|center|600px||]]</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*&#039;&#039;&#039;Temperaturprofil LightCycler&#039;&#039;&#039;</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*&#039;&#039;&#039;Temperaturprofil LightCycler&#039;&#039;&#039;</div></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>**
Enzymaktivering: 95 <del class="diffchange diffchange-inline">oC </del>10 min. Slope 20 <del class="diffchange diffchange-inline">oC</del>/s. 1 cykel.
</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>**
Enzymaktivering: 95 <ins class="diffchange diffchange-inline">°C </ins>10 min. Slope 20 <ins class="diffchange diffchange-inline">°C</ins>/s. 1 cykel.
</div></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>**Amplifiering: 95 <del class="diffchange diffchange-inline">oC </del>15 sek, 50 <del class="diffchange diffchange-inline">oC </del>15 sek, 72 <del class="diffchange diffchange-inline">oC </del>25 sek. 40 cykler. Slope 20 <del class="diffchange diffchange-inline">oC</del>/s.  </div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>**Amplifiering: 95 <ins class="diffchange diffchange-inline">°C </ins>15 sek, 50 <ins class="diffchange diffchange-inline">°C </ins>15 sek, 72 <ins class="diffchange diffchange-inline">°C </ins>25 sek. 40 cykler. Slope 20 <ins class="diffchange diffchange-inline">°C</ins>/s.  </div></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>**Kylning: 40 <del class="diffchange diffchange-inline">oC </del>60 sek. Slope 20 <del class="diffchange diffchange-inline">oC</del>/s.</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>**Kylning: 40 <ins class="diffchange diffchange-inline">°C </ins>60 sek. Slope 20 <ins class="diffchange diffchange-inline">°C</ins>/s.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>
Avläsning med color compensation. &#039;&#039;VanA&#039;&#039; och &#039;&#039;vanB&#039;&#039; avläses i olika kanaler.  </div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>
Avläsning med color compensation. &#039;&#039;VanA&#039;&#039; och &#039;&#039;vanB&#039;&#039; avläses i olika kanaler.  </div></td></tr> <tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l75" >Rad 75:</td> <td colspan="2" class="diff-lineno">Rad 75:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>   </div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>   </div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*&#039;&#039;&#039;Temperaturprofil för Rotor-Gene
&#039;&#039;&#039;</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*&#039;&#039;&#039;Temperaturprofil för Rotor-Gene
&#039;&#039;&#039;</div></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>**Enzymaktivering: 95 <del class="diffchange diffchange-inline">oC </del>i 3 min</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>**Enzymaktivering: 95 <ins class="diffchange diffchange-inline">°C </ins>i 3 min</div></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>**
Amplifiering: 95 <del class="diffchange diffchange-inline">oC </del>i 3 sek, 60 <del class="diffchange diffchange-inline">oC </del>i 20 sek. 40 cykler.
</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>**
Amplifiering: 95 <ins class="diffchange diffchange-inline">°C </ins>i 3 sek, 60 <ins class="diffchange diffchange-inline">°C </ins>i 20 sek. 40 cykler.
</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>&#039;&#039;VanA&#039;&#039; och &#039;&#039;vanB&#039;&#039; avläses i olika kanaler. Smältpunktanalys behövs inte.</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>&#039;&#039;VanA&#039;&#039; och &#039;&#039;vanB&#039;&#039; avläses i olika kanaler. Smältpunktanalys behövs inte.</div></td></tr> <tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l92" >Rad 92:</td> <td colspan="2" class="diff-lineno">Rad 92:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*&#039;&#039;&#039;Temperaturprofil LightCycler&#039;&#039;&#039;</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*&#039;&#039;&#039;Temperaturprofil LightCycler&#039;&#039;&#039;</div></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>**
Enzymaktivering: 95 <del class="diffchange diffchange-inline">oC </del>10 min. Slope 20 <del class="diffchange diffchange-inline">oC</del>/s. 1 cykel.</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>**
Enzymaktivering: 95 <ins class="diffchange diffchange-inline">°C </ins>10 min. Slope 20 <ins class="diffchange diffchange-inline">°C</ins>/s. 1 cykel.</div></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>**
Amplifiering: 95 <del class="diffchange diffchange-inline">oC </del>15 sek, 54 <del class="diffchange diffchange-inline">oC </del>10 sek, 72 <del class="diffchange diffchange-inline">oC </del>25 sek. 35 cykler. Slope 20 <del class="diffchange diffchange-inline">oC</del>/s.  </div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>**
Amplifiering: 95 <ins class="diffchange diffchange-inline">°C </ins>15 sek, 54 <ins class="diffchange diffchange-inline">°C </ins>10 sek, 72 <ins class="diffchange diffchange-inline">°C </ins>25 sek. 35 cykler. Slope 20 <ins class="diffchange diffchange-inline">°C</ins>/s.  </div></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>**Smältpunktsanalys: 60–90 <del class="diffchange diffchange-inline">oC</del>. Slope 0,1 <del class="diffchange diffchange-inline">oC</del>/s
</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>**Smältpunktsanalys: 60–90 <ins class="diffchange diffchange-inline">°C</ins>. Slope 0,1 <ins class="diffchange diffchange-inline">°C</ins>/s
</div></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>**Kylning: 40 <del class="diffchange diffchange-inline">oC </del>30 sek. Slope 20 <del class="diffchange diffchange-inline">oC</del>/s.</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>**Kylning: 40 <ins class="diffchange diffchange-inline">°C </ins>30 sek. Slope 20 <ins class="diffchange diffchange-inline">°C</ins>/s.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>Avläsning: Metoden är en cybergreenmetod och läses av i våglängd 530 nm. Positiva prov ska uppvisa en amplifieringskurva samt att produkten ska ha en smälttopp runt 84–85 <del class="diffchange diffchange-inline">oC</del>.</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Avläsning: Metoden är en cybergreenmetod och läses av i våglängd 530 nm. Positiva prov ska uppvisa en amplifieringskurva samt att produkten ska ha en smälttopp runt 84–85 <ins class="diffchange diffchange-inline">°C</ins>.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>[[Kategori:Laboratoriemetodik]]</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>[[Kategori:Laboratoriemetodik]]</div></td></tr> </table> Magnus Thore https://referensmetodik.folkhalsomyndigheten.se/index.php?title=Bilagor_till_VRE-diagnostiken&diff=8201&oldid=prev Magnus Thore: /* PCR-metod för påvisande av van-gener */ 2012-02-26T09:38:08Z <p><span dir="auto"><span class="autocomment">PCR-metod för påvisande av van-gener</span></span></p> <table class="diff diff-contentalign-left diff-editfont-monospace" data-mw="interface"> <col class="diff-marker" /> <col class="diff-content" /> <col class="diff-marker" /> <col class="diff-content" /> <tr class="diff-title" lang="sv"> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Äldre version</td> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Versionen från 26 februari 2012 kl. 09.38</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l88" >Rad 88:</td> <td colspan="2" class="diff-lineno">Rad 88:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">[[Fil:VerifieringEnterokocker.jpg|thumb|center|600px||]]</ins></div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>Temperaturprofil <del class="diffchange diffchange-inline">LightCycler
Enzymaktivering</del>: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
Amplifiering: 95 oC 15 sek, 54 oC 10 sek, 72 oC 25 sek. 35 cykler. Slope 20 oC/s. Smältpunktsanalys: 60–90 oC. Slope 0,1 oC/<del class="diffchange diffchange-inline">s
Kylning</del>: 40 oC 30 sek. Slope 20 oC/s.Avläsning: Metoden är en <del class="diffchange diffchange-inline">cybergreen metod </del>och läses av i våglängd 530 nm. Positiva prov ska uppvisa en amplifieringskurva samt att produkten ska ha en smälttopp runt 84–85 oC.<del class="diffchange diffchange-inline">62</del></div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins class="diffchange diffchange-inline">*&#039;&#039;&#039;</ins>Temperaturprofil <ins class="diffchange diffchange-inline">LightCycler&#039;&#039;&#039;</ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins class="diffchange diffchange-inline">**
Enzymaktivering</ins>: 95 oC 10 min. Slope 20 oC/s. 1 cykel.</div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins class="diffchange diffchange-inline">**</ins>
Amplifiering: 95 oC 15 sek, 54 oC 10 sek, 72 oC 25 sek. 35 cykler. Slope 20 oC/s.  </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins class="diffchange diffchange-inline">**</ins>Smältpunktsanalys: 60–90 oC. Slope 0,1 oC/<ins class="diffchange diffchange-inline">s
</ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins class="diffchange diffchange-inline">**Kylning</ins>: 40 oC 30 sek. Slope 20 oC/s.</div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Avläsning: Metoden är en <ins class="diffchange diffchange-inline">cybergreenmetod </ins>och läses av i våglängd 530 nm. Positiva prov ska uppvisa en amplifieringskurva samt att produkten ska ha en smälttopp runt 84–85 oC.</div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins class="diffchange diffchange-inline">[[Kategori:Laboratoriemetodik]]</ins></div></td></tr> </table> Magnus Thore https://referensmetodik.folkhalsomyndigheten.se/index.php?title=Bilagor_till_VRE-diagnostiken&diff=8200&oldid=prev Magnus Thore: /* PCR-metod för påvisande av van-gener */ 2012-02-26T09:35:05Z <p><span dir="auto"><span class="autocomment">PCR-metod för påvisande av van-gener</span></span></p> <table class="diff diff-contentalign-left diff-editfont-monospace" data-mw="interface"> <col class="diff-marker" /> <col class="diff-content" /> <col class="diff-marker" /> <col class="diff-content" /> <tr class="diff-title" lang="sv"> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Äldre version</td> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Versionen från 26 februari 2012 kl. 09.35</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l51" >Rad 51:</td> <td colspan="2" class="diff-lineno">Rad 51:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>====PCR-metod för påvisande av van-gener====</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>====PCR-metod för påvisande av van-gener====</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del class="diffchange diffchange-inline">*</del>&#039;&#039;&#039;&#039;&#039;
vanA&#039;&#039; och &#039;&#039;vanB&#039;&#039; PCR för LightCycler (FRET-hybridiseringsprobe)&#039;&#039;&#039;</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>&#039;&#039;&#039;&#039;&#039;
vanA&#039;&#039; och &#039;&#039;vanB&#039;&#039; PCR för LightCycler (FRET-hybridiseringsprobe)&#039;&#039;&#039;</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Reaktionsvolymen är 20 μL varav 5 μl är templat. PCR mixen innehåller 1xHYB- master (Roche Diagnostics), 2 mM MgCl2, 1 μM av varje primer och 0.2 μM av varje hybridiserings-probe.</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Reaktionsvolymen är 20 μL varav 5 μl är templat. PCR mixen innehåller 1xHYB- master (Roche Diagnostics), 2 mM MgCl2, 1 μM av varje primer och 0.2 μM av varje hybridiserings-probe.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div>[[Fil:VanAochvanB_LightCycler.jpg]]</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>[[Fil:VanAochvanB_LightCycler.jpg<ins class="diffchange diffchange-inline">|thumb|center|600px||</ins>]]</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*&#039;&#039;&#039;Temperaturprofil LightCycler&#039;&#039;&#039;</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>*&#039;&#039;&#039;Temperaturprofil LightCycler&#039;&#039;&#039;</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>**
Enzymaktivering: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>**
Enzymaktivering: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
</div></td></tr> <tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l66" >Rad 66:</td> <td colspan="2" class="diff-lineno">Rad 66:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Smältpunktanalys behövs inte.</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Smältpunktanalys behövs inte.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del class="diffchange diffchange-inline">*</del>&#039;&#039;&#039;&#039;&#039;VanA&#039;&#039; och &#039;&#039;vanB&#039;&#039; PCR för Rotor-Gene (TaqMan probe)&#039;&#039;&#039;</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins class="diffchange diffchange-inline">------------------------</ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>&#039;&#039;&#039;&#039;&#039;VanA&#039;&#039; och &#039;&#039;vanB&#039;&#039; PCR för Rotor-Gene (TaqMan probe)&#039;&#039;&#039;</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Reaktionsvolymen är 20 μL varav 2 μl är templat. PCR mixen innehåller 1x perfeCta qPCR FastMix (Quanta bioscience), 0,2 μM av varje primer, 0,075 μM av varje Taqman-probe.</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>Reaktionsvolymen är 20 μL varav 2 μl är templat. PCR mixen innehåller 1x perfeCta qPCR FastMix (Quanta bioscience), 0,2 μM av varje primer, 0,075 μM av varje Taqman-probe.</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">[[Fil:VanAochvanB_Rotor-Gene.jpg|thumb|center|600px||]]</ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;"> </ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">*&#039;&#039;&#039;Temperaturprofil för Rotor-Gene
&#039;&#039;&#039;</ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">**Enzymaktivering: 95 oC i 3 min</ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">**
Amplifiering: 95 oC i 3 sek, 60 oC i 20 sek. 40 cykler.
</ins></div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">&#039;&#039;VanA&#039;&#039; och &#039;&#039;vanB&#039;&#039; avläses i olika kanaler. Smältpunktanalys behövs inte.</ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">-----------------------------</ins></div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">&#039;&#039;&#039;PCR-metod för molekylärbiologisk verifiering av &#039;&#039;E. faecalis&#039;&#039; och &#039;&#039;E. faecium&#039;&#039; </ins></div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del class="diffchange diffchange-inline">Primers
vanA-F 5 ́-CGGCAAGACAATATGACAGCAA-3 ́ vanA-R 5 ́</del>-<del class="diffchange diffchange-inline">TCAGTACAATGCGGCCGTTA-3 ́ vanB-F 5 ́-GGGAGGATGGTGCGATACA-3 ́ vanB-R 5 ́-CCGAAA TCGCTTGCTCAA-3 ́Prober    vanA  5 ́HEX-CAGTTATAACCGTTCCCGCAGACCTT-BHQ1-3 ́    vanB  5 ́6FAM-CTTTGTGAAGCCGGCACGGTCAGGTT-BBQ-3 ́ Storlek på </del>PCR<del class="diffchange diffchange-inline">-fragmentet för vanA är 322 baspar och för vanB 329 baspar.61Temperaturprofil </del>för <del class="diffchange diffchange-inline">Rotor-Gene
Enzymaktivering</del>: <del class="diffchange diffchange-inline">95 oC i 3 min
Amplifiering: 95 oC i 3 sek, 60 oC i 20 sek. 40 cykler.
VanA och vanB avläses i olika kanaler. Smältpunktanalys behövs inte.PCR-metod för molekylärbiologisk verifiering av E. faecalis och E. faecium DdlE PCR för LightCyclerReaktionsvolymen </del>är 20 μL varav 4 μl är templat. PCR mixen innehåller 1x LC- master, 4 mM MgCl2, 0,5 μM av varje primer och 0,04 units/μL FastStart Taq DNA-polymeras.LC-master innehåller 2x FastStart PCR-buffert (Roche diagnostics), 4 % DMSO (Sigma), 0,4 mM dNTP (Roche diagnostics), 0,2 mg/mL BSA (Roche diagnostics) och 1,2x SybrGreen I (Molecular Probes).<del class="diffchange diffchange-inline">Primer
EFS-F 5 ́-CACCTGAAGAAACAGGC-3 ́ EFS-R 5 ́-ATGGCTACTTCAATTTCACG-3 ́EFM-F 5 ́-AATAATAAAATCGAAATGCAGATTCC-3 ́ EFM-R 5 ́-TCCAGCCTCTTCTTTATTTCTCTTTAT-3 ́Storleken på DNA-fragmentet för E. faecalis är 475 baspar och för E. faecium 335 baspar.</del>Temperaturprofil LightCycler
Enzymaktivering: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
Amplifiering: 95 oC 15 sek, 54 oC 10 sek, 72 oC 25 sek. 35 cykler. Slope 20 oC/s. Smältpunktsanalys: 60–90 oC. Slope 0,1 oC/s
Kylning: 40 oC 30 sek. Slope 20 oC/s.Avläsning: Metoden är en cybergreen metod och läses av i våglängd 530 nm. Positiva prov ska uppvisa en amplifieringskurva samt att produkten ska ha en smälttopp runt 84–85 oC.62</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins class="diffchange diffchange-inline">&#039;&#039;&#039;&#039;&#039;DdlE&#039;&#039;</ins>-PCR för <ins class="diffchange diffchange-inline">LightCycler&#039;&#039;&#039;</ins>: <ins class="diffchange diffchange-inline">Reaktionsvolymen </ins>är 20 μL varav 4 μl är templat. PCR mixen innehåller 1x LC- master, 4 mM MgCl2, 0,5 μM av varje primer och 0,04 units/μL FastStart Taq DNA-polymeras.</div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>LC-master innehåller 2x FastStart PCR-buffert (Roche diagnostics), 4 % DMSO (Sigma), 0,4 mM dNTP (Roche diagnostics), 0,2 mg/mL BSA (Roche diagnostics) och 1,2x SybrGreen I (Molecular Probes).</div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Temperaturprofil LightCycler
Enzymaktivering: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
Amplifiering: 95 oC 15 sek, 54 oC 10 sek, 72 oC 25 sek. 35 cykler. Slope 20 oC/s. Smältpunktsanalys: 60–90 oC. Slope 0,1 oC/s
Kylning: 40 oC 30 sek. Slope 20 oC/s.Avläsning: Metoden är en cybergreen metod och läses av i våglängd 530 nm. Positiva prov ska uppvisa en amplifieringskurva samt att produkten ska ha en smälttopp runt 84–85 oC.62</div></td></tr> </table> Magnus Thore https://referensmetodik.folkhalsomyndigheten.se/index.php?title=Bilagor_till_VRE-diagnostiken&diff=8199&oldid=prev Magnus Thore: /* PCR-metod för påvisande av van-gener */ 2012-02-26T09:26:25Z <p><span dir="auto"><span class="autocomment">PCR-metod för påvisande av van-gener</span></span></p> <table class="diff diff-contentalign-left diff-editfont-monospace" data-mw="interface"> <col class="diff-marker" /> <col class="diff-content" /> <col class="diff-marker" /> <col class="diff-content" /> <tr class="diff-title" lang="sv"> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Äldre version</td> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Versionen från 26 februari 2012 kl. 09.26</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l56" >Rad 56:</td> <td colspan="2" class="diff-lineno">Rad 56:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">[[Fil:VanAochvanB_LightCycler.jpg]]</ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">*&#039;&#039;&#039;Temperaturprofil LightCycler&#039;&#039;&#039;</ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">**
Enzymaktivering: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
</ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">**Amplifiering: 95 oC 15 sek, 50 oC 15 sek, 72 oC 25 sek. 40 cykler. Slope 20 oC/s. </ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins style="font-weight: bold; text-decoration: none;">**Kylning: 40 oC 60 sek. Slope 20 oC/s.</ins></div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del class="diffchange diffchange-inline">Primers
vanA-F 5 ́-TCGGCAAGACAATATGACAG-3 ́ vanA-R 5 ́-GTACAATGCGGCCGTTA-3 ́ vanB-F 5 ́-GAGGATGGTGCGATACA-3 ́ vanB-R 5 ́-GATGGATGCGGAAGATA-3 ́Prober
vanA-FL 5 ́-ACGCAGTTATAACCGTTCCCG -FL-3 ́
vanA-705 5 ́-LC705-AGACCTTTCAGCAGAGGAGCG-PH-3 ́ vanB-FL 5 ́-CCTGTCTTTGTGAAGCCG-FL-3 ́
vanB-640 5 ́-LC640-CACGGTCAGGTTCGTCCTTTGGCGT-PH-3 ́Storlek på PCR-fragmentet för vanA är 322 baspar och för vanB 423 baspar.Temperaturprofil LightCycler
Enzymaktivering: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
Amplifiering: 95 oC 15 sek, 50 oC 15 sek, 72 oC 25 sek. 40 cykler. Slope 20 oC/s. Kylning: 40 oC 60 sek. Slope 20 oC/s.</del>
Avläsning med color compensation. VanA och vanB avläses i olika kanaler. Smältpunktanalys behövs inte.VanA och vanB PCR för Rotor-Gene (TaqMan probe)Reaktionsvolymen är 20 μL varav 2 μl är templat. PCR mixen innehåller 1x perfeCta qPCR FastMix (Quanta bioscience), 0,2 μM av varje primer, 0,075 μM av varje Taqman-probe.Primers
vanA-F 5 ́-CGGCAAGACAATATGACAGCAA-3 ́ vanA-R 5 ́-TCAGTACAATGCGGCCGTTA-3 ́ vanB-F 5 ́-GGGAGGATGGTGCGATACA-3 ́ vanB-R 5 ́-CCGAAA TCGCTTGCTCAA-3 ́Prober    vanA  5 ́HEX-CAGTTATAACCGTTCCCGCAGACCTT-BHQ1-3 ́    vanB  5 ́6FAM-CTTTGTGAAGCCGGCACGGTCAGGTT-BBQ-3 ́ Storlek på PCR-fragmentet för vanA är 322 baspar och för vanB 329 baspar.61Temperaturprofil för Rotor-Gene
Enzymaktivering: 95 oC i 3 min
Amplifiering: 95 oC i 3 sek, 60 oC i 20 sek. 40 cykler.
VanA och vanB avläses i olika kanaler. Smältpunktanalys behövs inte.PCR-metod för molekylärbiologisk verifiering av E. faecalis och E. faecium DdlE PCR för LightCyclerReaktionsvolymen är 20 μL varav 4 μl är templat. PCR mixen innehåller 1x LC- master, 4 mM MgCl2, 0,5 μM av varje primer och 0,04 units/μL FastStart Taq DNA-polymeras.LC-master innehåller 2x FastStart PCR-buffert (Roche diagnostics), 4 % DMSO (Sigma), 0,4 mM dNTP (Roche diagnostics), 0,2 mg/mL BSA (Roche diagnostics) och 1,2x SybrGreen I (Molecular Probes).Primer
EFS-F 5 ́-CACCTGAAGAAACAGGC-3 ́ EFS-R 5 ́-ATGGCTACTTCAATTTCACG-3 ́EFM-F 5 ́-AATAATAAAATCGAAATGCAGATTCC-3 ́ EFM-R 5 ́-TCCAGCCTCTTCTTTATTTCTCTTTAT-3 ́Storleken på DNA-fragmentet för E. faecalis är 475 baspar och för E. faecium 335 baspar.Temperaturprofil LightCycler
Enzymaktivering: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
Amplifiering: 95 oC 15 sek, 54 oC 10 sek, 72 oC 25 sek. 35 cykler. Slope 20 oC/s. Smältpunktsanalys: 60–90 oC. Slope 0,1 oC/s
Kylning: 40 oC 30 sek. Slope 20 oC/s.Avläsning: Metoden är en cybergreen metod och läses av i våglängd 530 nm. Positiva prov ska uppvisa en amplifieringskurva samt att produkten ska ha en smälttopp runt 84–85 oC.62</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>
Avläsning med color compensation. <ins class="diffchange diffchange-inline">&#039;&#039;</ins>VanA<ins class="diffchange diffchange-inline">&#039;&#039; </ins>och <ins class="diffchange diffchange-inline">&#039;&#039;</ins>vanB<ins class="diffchange diffchange-inline">&#039;&#039; </ins>avläses i olika kanaler.  </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Smältpunktanalys behövs inte.</div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins class="diffchange diffchange-inline">*&#039;&#039;&#039;&#039;&#039;</ins>VanA<ins class="diffchange diffchange-inline">&#039;&#039; </ins>och <ins class="diffchange diffchange-inline">&#039;&#039;</ins>vanB<ins class="diffchange diffchange-inline">&#039;&#039; </ins>PCR för Rotor-Gene (TaqMan probe)<ins class="diffchange diffchange-inline">&#039;&#039;&#039;</ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Reaktionsvolymen är 20 μL varav 2 μl är templat. PCR mixen innehåller 1x perfeCta qPCR FastMix (Quanta bioscience), 0,2 μM av varje primer, 0,075 μM av varje Taqman-probe.</div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Primers
vanA-F 5 ́-CGGCAAGACAATATGACAGCAA-3 ́ vanA-R 5 ́-TCAGTACAATGCGGCCGTTA-3 ́ vanB-F 5 ́-GGGAGGATGGTGCGATACA-3 ́ vanB-R 5 ́-CCGAAA TCGCTTGCTCAA-3 ́Prober    vanA  5 ́HEX-CAGTTATAACCGTTCCCGCAGACCTT-BHQ1-3 ́    vanB  5 ́6FAM-CTTTGTGAAGCCGGCACGGTCAGGTT-BBQ-3 ́ Storlek på PCR-fragmentet för vanA är 322 baspar och för vanB 329 baspar.61Temperaturprofil för Rotor-Gene
Enzymaktivering: 95 oC i 3 min
Amplifiering: 95 oC i 3 sek, 60 oC i 20 sek. 40 cykler.
VanA och vanB avläses i olika kanaler. Smältpunktanalys behövs inte.PCR-metod för molekylärbiologisk verifiering av E. faecalis och E. faecium DdlE PCR för LightCyclerReaktionsvolymen är 20 μL varav 4 μl är templat. PCR mixen innehåller 1x LC- master, 4 mM MgCl2, 0,5 μM av varje primer och 0,04 units/μL FastStart Taq DNA-polymeras.LC-master innehåller 2x FastStart PCR-buffert (Roche diagnostics), 4 % DMSO (Sigma), 0,4 mM dNTP (Roche diagnostics), 0,2 mg/mL BSA (Roche diagnostics) och 1,2x SybrGreen I (Molecular Probes).Primer
EFS-F 5 ́-CACCTGAAGAAACAGGC-3 ́ EFS-R 5 ́-ATGGCTACTTCAATTTCACG-3 ́EFM-F 5 ́-AATAATAAAATCGAAATGCAGATTCC-3 ́ EFM-R 5 ́-TCCAGCCTCTTCTTTATTTCTCTTTAT-3 ́Storleken på DNA-fragmentet för E. faecalis är 475 baspar och för E. faecium 335 baspar.Temperaturprofil LightCycler
Enzymaktivering: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
Amplifiering: 95 oC 15 sek, 54 oC 10 sek, 72 oC 25 sek. 35 cykler. Slope 20 oC/s. Smältpunktsanalys: 60–90 oC. Slope 0,1 oC/s
Kylning: 40 oC 30 sek. Slope 20 oC/s.Avläsning: Metoden är en cybergreen metod och läses av i våglängd 530 nm. Positiva prov ska uppvisa en amplifieringskurva samt att produkten ska ha en smälttopp runt 84–85 oC.62</div></td></tr> </table> Magnus Thore https://referensmetodik.folkhalsomyndigheten.se/index.php?title=Bilagor_till_VRE-diagnostiken&diff=8195&oldid=prev Magnus Thore den 26 februari 2012 kl. 09.20 2012-02-26T09:20:56Z <p></p> <table class="diff diff-contentalign-left diff-editfont-monospace" data-mw="interface"> <col class="diff-marker" /> <col class="diff-content" /> <col class="diff-marker" /> <col class="diff-content" /> <tr class="diff-title" lang="sv"> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">← Äldre version</td> <td colspan="2" style="background-color: #fff; color: #202122; text-align: center;">Versionen från 26 februari 2012 kl. 09.20</td> </tr><tr><td colspan="2" class="diff-lineno" id="mw-diff-left-l51" >Rad 51:</td> <td colspan="2" class="diff-lineno">Rad 51:</td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>====PCR-metod för påvisande av van-gener====</div></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"><div>====PCR-metod för påvisande av van-gener====</div></td></tr> <tr><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td><td class='diff-marker'> </td><td style="background-color: #f8f9fa; color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #eaecf0; vertical-align: top; white-space: pre-wrap;"></td></tr> <tr><td class='diff-marker'>−</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #ffe49c; vertical-align: top; white-space: pre-wrap;"><div><del class="diffchange diffchange-inline"> </del>
vanA och vanB PCR för LightCycler (FRET-hybridiseringsprobe)Reaktionsvolymen är 20 μL varav 5 μl är templat. PCR mixen innehåller 1xHYB- master (Roche Diagnostics), 2 mM MgCl2, 1 μM av varje primer och 0.2 μM av varje hybridiserings-probe.Primers
vanA-F 5 ́-TCGGCAAGACAATATGACAG-3 ́ vanA-R 5 ́-GTACAATGCGGCCGTTA-3 ́ vanB-F 5 ́-GAGGATGGTGCGATACA-3 ́ vanB-R 5 ́-GATGGATGCGGAAGATA-3 ́Prober
vanA-FL 5 ́-ACGCAGTTATAACCGTTCCCG -FL-3 ́
vanA-705 5 ́-LC705-AGACCTTTCAGCAGAGGAGCG-PH-3 ́ vanB-FL 5 ́-CCTGTCTTTGTGAAGCCG-FL-3 ́
vanB-640 5 ́-LC640-CACGGTCAGGTTCGTCCTTTGGCGT-PH-3 ́Storlek på PCR-fragmentet för vanA är 322 baspar och för vanB 423 baspar.Temperaturprofil LightCycler
Enzymaktivering: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
Amplifiering: 95 oC 15 sek, 50 oC 15 sek, 72 oC 25 sek. 40 cykler. Slope 20 oC/s. Kylning: 40 oC 60 sek. Slope 20 oC/s.
Avläsning med color compensation. VanA och vanB avläses i olika kanaler. Smältpunktanalys behövs inte.VanA och vanB PCR för Rotor-Gene (TaqMan probe)Reaktionsvolymen är 20 μL varav 2 μl är templat. PCR mixen innehåller 1x perfeCta qPCR FastMix (Quanta bioscience), 0,2 μM av varje primer, 0,075 μM av varje Taqman-probe.Primers
vanA-F 5 ́-CGGCAAGACAATATGACAGCAA-3 ́ vanA-R 5 ́-TCAGTACAATGCGGCCGTTA-3 ́ vanB-F 5 ́-GGGAGGATGGTGCGATACA-3 ́ vanB-R 5 ́-CCGAAA TCGCTTGCTCAA-3 ́Prober    vanA  5 ́HEX-CAGTTATAACCGTTCCCGCAGACCTT-BHQ1-3 ́    vanB  5 ́6FAM-CTTTGTGAAGCCGGCACGGTCAGGTT-BBQ-3 ́ Storlek på PCR-fragmentet för vanA är 322 baspar och för vanB 329 baspar.61Temperaturprofil för Rotor-Gene
Enzymaktivering: 95 oC i 3 min
Amplifiering: 95 oC i 3 sek, 60 oC i 20 sek. 40 cykler.
VanA och vanB avläses i olika kanaler. Smältpunktanalys behövs inte.PCR-metod för molekylärbiologisk verifiering av E. faecalis och E. faecium DdlE PCR för LightCyclerReaktionsvolymen är 20 μL varav 4 μl är templat. PCR mixen innehåller 1x LC- master, 4 mM MgCl2, 0,5 μM av varje primer och 0,04 units/μL FastStart Taq DNA-polymeras.LC-master innehåller 2x FastStart PCR-buffert (Roche diagnostics), 4 % DMSO (Sigma), 0,4 mM dNTP (Roche diagnostics), 0,2 mg/mL BSA (Roche diagnostics) och 1,2x SybrGreen I (Molecular Probes).Primer
EFS-F 5 ́-CACCTGAAGAAACAGGC-3 ́ EFS-R 5 ́-ATGGCTACTTCAATTTCACG-3 ́EFM-F 5 ́-AATAATAAAATCGAAATGCAGATTCC-3 ́ EFM-R 5 ́-TCCAGCCTCTTCTTTATTTCTCTTTAT-3 ́Storleken på DNA-fragmentet för E. faecalis är 475 baspar och för E. faecium 335 baspar.Temperaturprofil LightCycler
Enzymaktivering: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
Amplifiering: 95 oC 15 sek, 54 oC 10 sek, 72 oC 25 sek. 35 cykler. Slope 20 oC/s. Smältpunktsanalys: 60–90 oC. Slope 0,1 oC/s
Kylning: 40 oC 30 sek. Slope 20 oC/s.Avläsning: Metoden är en cybergreen metod och läses av i våglängd 530 nm. Positiva prov ska uppvisa en amplifieringskurva samt att produkten ska ha en smälttopp runt 84–85 oC.62</div></td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div><ins class="diffchange diffchange-inline">*&#039;&#039;&#039;&#039;&#039;</ins>
vanA<ins class="diffchange diffchange-inline">&#039;&#039; </ins>och <ins class="diffchange diffchange-inline">&#039;&#039;</ins>vanB<ins class="diffchange diffchange-inline">&#039;&#039; </ins>PCR för LightCycler (FRET-hybridiseringsprobe)<ins class="diffchange diffchange-inline">&#039;&#039;&#039;</ins></div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Reaktionsvolymen är 20 μL varav 5 μl är templat. PCR mixen innehåller 1xHYB- master (Roche Diagnostics), 2 mM MgCl2, 1 μM av varje primer och 0.2 μM av varje hybridiserings-probe.</div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div> </div></td></tr> <tr><td colspan="2"> </td><td class='diff-marker'>+</td><td style="color: #202122; font-size: 88%; border-style: solid; border-width: 1px 1px 1px 4px; border-radius: 0.33em; border-color: #a3d3ff; vertical-align: top; white-space: pre-wrap;"><div>Primers
vanA-F 5 ́-TCGGCAAGACAATATGACAG-3 ́ vanA-R 5 ́-GTACAATGCGGCCGTTA-3 ́ vanB-F 5 ́-GAGGATGGTGCGATACA-3 ́ vanB-R 5 ́-GATGGATGCGGAAGATA-3 ́Prober
vanA-FL 5 ́-ACGCAGTTATAACCGTTCCCG -FL-3 ́
vanA-705 5 ́-LC705-AGACCTTTCAGCAGAGGAGCG-PH-3 ́ vanB-FL 5 ́-CCTGTCTTTGTGAAGCCG-FL-3 ́
vanB-640 5 ́-LC640-CACGGTCAGGTTCGTCCTTTGGCGT-PH-3 ́Storlek på PCR-fragmentet för vanA är 322 baspar och för vanB 423 baspar.Temperaturprofil LightCycler
Enzymaktivering: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
Amplifiering: 95 oC 15 sek, 50 oC 15 sek, 72 oC 25 sek. 40 cykler. Slope 20 oC/s. Kylning: 40 oC 60 sek. Slope 20 oC/s.
Avläsning med color compensation. VanA och vanB avläses i olika kanaler. Smältpunktanalys behövs inte.VanA och vanB PCR för Rotor-Gene (TaqMan probe)Reaktionsvolymen är 20 μL varav 2 μl är templat. PCR mixen innehåller 1x perfeCta qPCR FastMix (Quanta bioscience), 0,2 μM av varje primer, 0,075 μM av varje Taqman-probe.Primers
vanA-F 5 ́-CGGCAAGACAATATGACAGCAA-3 ́ vanA-R 5 ́-TCAGTACAATGCGGCCGTTA-3 ́ vanB-F 5 ́-GGGAGGATGGTGCGATACA-3 ́ vanB-R 5 ́-CCGAAA TCGCTTGCTCAA-3 ́Prober    vanA  5 ́HEX-CAGTTATAACCGTTCCCGCAGACCTT-BHQ1-3 ́    vanB  5 ́6FAM-CTTTGTGAAGCCGGCACGGTCAGGTT-BBQ-3 ́ Storlek på PCR-fragmentet för vanA är 322 baspar och för vanB 329 baspar.61Temperaturprofil för Rotor-Gene
Enzymaktivering: 95 oC i 3 min
Amplifiering: 95 oC i 3 sek, 60 oC i 20 sek. 40 cykler.
VanA och vanB avläses i olika kanaler. Smältpunktanalys behövs inte.PCR-metod för molekylärbiologisk verifiering av E. faecalis och E. faecium DdlE PCR för LightCyclerReaktionsvolymen är 20 μL varav 4 μl är templat. PCR mixen innehåller 1x LC- master, 4 mM MgCl2, 0,5 μM av varje primer och 0,04 units/μL FastStart Taq DNA-polymeras.LC-master innehåller 2x FastStart PCR-buffert (Roche diagnostics), 4 % DMSO (Sigma), 0,4 mM dNTP (Roche diagnostics), 0,2 mg/mL BSA (Roche diagnostics) och 1,2x SybrGreen I (Molecular Probes).Primer
EFS-F 5 ́-CACCTGAAGAAACAGGC-3 ́ EFS-R 5 ́-ATGGCTACTTCAATTTCACG-3 ́EFM-F 5 ́-AATAATAAAATCGAAATGCAGATTCC-3 ́ EFM-R 5 ́-TCCAGCCTCTTCTTTATTTCTCTTTAT-3 ́Storleken på DNA-fragmentet för E. faecalis är 475 baspar och för E. faecium 335 baspar.Temperaturprofil LightCycler
Enzymaktivering: 95 oC 10 min. Slope 20 oC/s. 1 cykel.
Amplifiering: 95 oC 15 sek, 54 oC 10 sek, 72 oC 25 sek. 35 cykler. Slope 20 oC/s. Smältpunktsanalys: 60–90 oC. Slope 0,1 oC/s
Kylning: 40 oC 30 sek. Slope 20 oC/s.Avläsning: Metoden är en cybergreen metod och läses av i våglängd 530 nm. Positiva prov ska uppvisa en amplifieringskurva samt att produkten ska ha en smälttopp runt 84–85 oC.62</div></td></tr> </table> Magnus Thore