Detection and quantitation of bovine respiratory syncytial virus using real-time quantitative RT-PCR and quantitative competitive RT-PCR assays

Detection and quantitation of bovine respiratory syncytial virus using real-time quantitative RT-PCR and quantitative competitive RT-PCR assays.

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 bovi...

Comparison of four RT-PCR assays for detection of bovine respiratory syncytial virus.

RT-PCR assays for detection of BRSV, based on four different sets of primers were optimized and evaluated for their sensitivity and specificity. Primers used in this study were specific for genes encoding three BRSV proteins, nucleoprotein N and glycoproteins F and G. Our results indicated that RT-PCR with primers B7:B8 for G protein was the most efficient in detecting BRSV. Starters B7:B8 reac...

Quantitative RT-PCR using a PCR-generated competitive internal standard.

One approach to quantitate RNA is the use of a known number of competitive internal standard molecules that are reverse-transcribed and subsequently co-amplified under the same reaction conditions as the target sequence (3). This technique, called competitive reverse transcription polymerase chain reaction (cRT-PCR) has been applied to monitor the level of hepatitis C virus (HCV) RNA in patient...

Quantitative PCR and RT-PCR in virology.

I n the last few years, molecular hybridization methods have been used extensively in basic and applied virology because of their technical flexibility and high specificity. Using these techniques, the detection of DNA and RNA viruses directly from clinical specimens, the analysis of the specific transcriptional activity of viral genes in vitro and in vivo, and the study of virus-host relations...

Real-time quantitative RT-PCR: design, calculations, and statistics.