Loading...
Please wait, while we are loading the content...
Detection and quantitation of bovine respiratory syncytial virus using real-time quantitative RT-PCR and quantitative competitive RT-PCR assays.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Achenbach, Jenna E. Topliff, Christina L. Vassilev, Ventzislav B. Donis, Ruben O. Eskridge, Kent M. Kelling, Clayton L. |
| Copyright Year | 2004 |
| Abstract | A single tube, fluorogenic probe-based, real-time quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) assay was developed for detection and quantitation of bovine respiratory syncytial virus (BRSV) using BioRad's iCycler iQ. Real-time Q-RT-PCR was compared with quantitative competitive RT-PCR (QC-RT-PCR) and viral titers. Viral mRNA levels were measured in BRSV-infected bovine turbinate cell lysate harvested at eight time points (1.5, 6, 12, 24, 36, 48, 60, 72 h) post-infection. A homologous BRSV cRNA standard was used for quantitation of the mRNA by plotting a standard curve of cycle threshold (Ct) values versus standard 10-fold dilutions of cRNA of known concentrations. Detection as low as 171 copies/microl of standard BRSV cRNA was possible. For QC-RT-PCR, a competitor RNA molecule having a deletion was designed and used for quantitation of the BRSV viral mRNA. The results of real-time Q-RT-PCR and QC-RT-PCR assays showed a positive correlation. Real-time Q-RT-PCR was a sensitive, specific, rapid, and efficient method that eliminates the post-PCR processing steps when compared to QC-RT-PCR. Quantitation of BRSV using real-time Q-RT-PCR will have application in studies aimed at understanding the pathogenesis of BRSV. |
| Starting Page | 1 |
| Ending Page | 6 |
| Page Count | 6 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | http://digitalcommons.unl.edu/cgi/viewcontent.cgi?article=1109&context=virologypub |
| Alternate Webpage(s) | http://digitalcommons.unl.edu/cgi/viewcontent.cgi?amp%3Bcontext=virologypub&article=1109 |
| PubMed reference number | 15350726v1 |
| Volume Number | 121 |
| Issue Number | 1 |
| Journal | Journal of virological methods |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Cattle Citric Acid Cycle Complementary RNA Copy (object) Deletion Mutation Lysate Murine sarcoma viruses Nasal turbinate bone structure Quantitation Respiratory Syncytial Virus, Bovine Respiratory syncytial virus Reverse Transcriptase Polymerase Chain Reaction Reverse Transcription |
| Content Type | Text |
| Resource Type | Article |
