Quantitation of RNA coliphage inactivated by UV radiation using RT-PCR method

Hiroyuki KATAYAMA, Masahiro OTAKI, Shinichiro OHGAKI
1997 Environmental Engineering Research  
PCR (polymerase chain reaction) is a useful method for viral monitoring because of its high sensitivity and short time detection. However, disadvantages of PCR method for viral monitoring are pointed out to be lack of quantification and no differentiation of infectious viruses from noninfectious viruses. MPN method using RT(reverse transcription)-PCR was applied for enumeration of RNA coliphage QƒÀ, and nested PCR was applied to promote the sensitivity of RT-PCR. This method gave consistently
more » ... e same order of values with plaque assay with double agar layer (host: E.coll K12 F+ A/ƒÉ). The authors developed"long RT-short PCR method", which demands a longer RT region to be detected. The RT region was 1909bp, while total RNA of QƒÀ is 4217bp. This method was also applied to QƒÀ disinfected by UV radiation which had no plaque forming ability. The results gave the same value with the MPN value of QƒÀ sample which was not inactivated with UV radiation. These results were analyzed with one hit model.
doi:10.11532/proes1992.34.83 fatcat:zl2jxzbujzf6tbtfwdspnytzam