None of the rRT-PCR or cRT-PCR assays cross-reacted with any of the other common equine respiratory viruses. With the exception of one cRT-PCR assay, the detection limit of all of these assays was 1 plaque forming unit per ml (pfu/ml). Conclusion The newly developed rRT-PCR and cRT-...
Further, nested PCR has been carried out using p1/p7 and fU5/rU3 primers and resulted in the amplification product size of 890bp. From this amplified product, specifically a target of 69bp from the 16S rRNA gene region has been detected through primers conjugated with Taqman probe in a ...
All RT-PCRs were run on the CFX 96 real-time PCR cycler (Bio-Rad, Hercules, CA, USA) with the AgPath-ID™ One-Step RT-PCR Reagents of Applied Biosystems™ (Waltham, USA). The temperature profile used for BlueTYPE and all individual RT-qPCR runs was 10 min at 45 °C (rev...
Cross-contamination of samples especially during the RNA extraction process is a real concern with any RT-PCR reaction, but it is recognized that precautions should be taken to limit contamination at every step of the procedure. The viral load in the trachea of birds given different dosages of...
None of the rRT-PCR or cRT-PCR assays cross-reacted with any of the other common equine respiratory viruses. With the exception of one cRT-PCR assay, the detection limit of all of these assays was 1 plaque forming unit per ml (pfu/ml). Conclusion The newly developed rRT-PCR and cRT-...
Results We have developed and evaluated a multiplex one-tube RT-PCR using a combination of GI and GII specific primers and probes located in the junction between ORF1 and ORF2 of the norovirus genome. Comparable results for sensitivity and specificity to those from our in-house nested PCR ...