Use of bacteriophage MS2 as an internal control in viral reverse transcription-PCR assays.

ABSTRACT: Diagnostic systems based on reverse transcription (RT)-PCR are widely used for the detection of viral genomes in different human specimens. The application of internal controls (IC) to monitor each step of nucleic acid amplification is necessary to prevent false-negative results due to inhibition or human error. In this study, we designed various real-time RT-PCRs utilizing the coliphage MS2 replicase gene, which differ in detection format, amplicon size, and efficiency of amplification. These noncompetitive IC assays, using TaqMan, hybridization probe, or duplex scorpion probe techniques, were tested on the LightCycler and Rotorgene systems. In our approach, clinical specimens were spiked with the control virus to monitor the efficiency of extraction, reverse transcription, and amplificat

SUBMITTER: Dreier J 

PROVIDER: S-EPMC1234060 | biostudies-literature | 2005 Sep

REPOSITORIES: biostudies-literature