Application of two-step quantitative reverse-transcription PCR to bacterial diagnostics.
Use of high copy number bacterial RNA offers several advantages in a diagnostics context compared with current deoxyribonucleic acid-based assays. The opportunity to only detect viable cells by targeting labile RNA transcripts may create an opportunity for "real-time" monitoring of pathogen load in response to a treatment regimen, while the natural amplification provided by the relative abundance of the RNA target compared with its corresponding gene opens a door to potential nonamplified direct detection technologies. In this chapter, a method is described to accurately quantify specific RNA transcripts and thus determine their potential utility as "high-copy" targets. The quantification method described also has application in gene-expression analysis.
Glynn
Barry
B
National Diagnostics Centre, National University of Ireland, Galway.
D001419:Bacteria / Q000145:classification* / Q000235:genetics; D018628:Gene Dosage; D012329:RNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D018075:RNA, Complementary / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D012329:RNA, Bacterial; D018075:RNA, Complementary