A one step quantitative RT-PCR for detection of SARS coronavirus with an internal control for PCR inhibitors

One-Step Reverse Transcription PCR (RT-PCR). Fatemeh Mehrpooyan*

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...

Development of a Multiplex RT-PCR Assay for Detection of the Causal Agents of Citrus Tristeza and Cachexia Diseases with Coamplification of Plant mRNA as an Internal Control

Background and Aims: Plant certification programs need reliable, fast, cheap and sensitive methods for detection of systemic pathogens with special interest in virus and viroid detection. Reverse transcriptase-polymerase chain reaction (RT-PCR) has been documented as an alternative assay for certification of plant propagating materials. The main object of the present study was the optimization ...

Rapid one-step quantitative reverse transcriptase PCR assay with competitive internal positive control for detection of enteroviruses in environmental samples.

Human enteroviruses can serve as a more accurate indicator of human fecal contamination than conventional bacteriological fecal indicators. We describe here a quantitative reverse transcriptase PCR (qRT-PCR) assay specifically tailored to detect these viruses in environmental waters. The assay included a competitive internal positive control (CIPC) that allowed the inhibition of qRT-PCRs to be ...

Guidelines for quantitative rt-PCR.

Something I notice often in supervising new graduate students is their frustration with the irreproducibility of their biological data. For example, I am asked frequently ‘‘how many biological replicates do I need for qRT-PCR’’? The answer, of course, lies first with the students themselves. They need as many replicates as will persuade them of the validity of their observations. However, for m...