A one-tube nucleic acid extraction and amplification (ERTPCR) method for detecting RNA viruses

Extraction Reverse Transcription Polymerase Chain Reaction (ERTPCR), a rapi d and simple method which allows detection of RNA viruses, is described. Un like conventional Reverse Transcription Polymerase Chain Reaction(RTPCR), w hich requires extraction of viral nucleic acid prior to reverse transcripti on and cDNA amplification, ERTPCR allows these procedures to be accomplishe d in one step in the same reaction tube. This is facilitated through heat e xtraction of viral RNA, and the use of a thermostable enzyme which can perf orm reverse transcription and cDNA amplification in the same buffer system. The technique is at least as sensitive as a standard RTPCR. Its use in det ecting rotavirus in feces, and hepatitis A virus in infected tissue culture s, is demonstrated. A silica spin column was used to remove RTPCR inhibitor s from these samples.