Key result
Extraction Reverse Transcription Polymerase Chain Reaction (ERTPCR) was at least as sensitive as standard RTPCR for detecting RNA viruses in a single reaction tube.
ERTPCR provides a rapid, one-tube method for RNA virus detection that is as sensitive as standard RTPCR.
May simplify RNA virus detection; animal data leave clinical translation open.
Extraction Reverse Transcription Polymerase Chain Reaction (ERTPCR), a rapid and simple method which allows detection of RNA viruses, is described. Unlike conventional Reverse Transcription Polymerase Chain Reaction(RTPCR), which requires extraction of viral nucleic acid prior to reverse transcription and cDNA amplification, ERTPCR allows these procedures to be accomplished in one step in the same reaction tube. This is facilitated through heat extraction of viral RNA, and the use of a thermostable enzyme which can perform reverse transcription and cDNA amplification in the same buffer system. The technique is at least as sensitive as a standard RTPCR. Its use in detecting rotavirus in feces, and hepatitis A virus in infected tissue cultures, is demonstrated. A silica spin column was used to remove RTPCR inhibitors from these samples.
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Cook et al. (1999) studied RNA viruses. Extraction Reverse Transcription Polymerase Chain Reaction (ERTPCR) vs. Standard RTPCR was evaluated on Sensitivity of viral RNA detection. Extraction Reverse Transcription Polymerase Chain Reaction (ERTPCR) was at least as sensitive as standard RTPCR for detecting RNA viruses in a single reaction tube.
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