Detection method of viral pollution in environmental water

摘要

PURPOSE: Provided is a detection method of Adenovirus and Enteric virus contamination from environment water by multiplex nested polymerase chain reaction PCR) analysis. This detection method can be applied to the detection of other generalized virus. CONSTITUTION: The detection method comprises: making two pairs of Adenovirus primers of 75nM-130nM, which are AV1, AV2, AV3 and AV4, containing SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 and SEQ ID NO: 7 respectively and three enteric virus primer of 200nM-300nM, which are EV1, EV2 and EV3, containing SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3; carrying on the first PCR by a) substituting enteric virus RNA to DNA by reverse transcription PCR between enteric virus primer EV2 and reverse transcriptase, b) adding Adenovirus primer AV1/AV2 and enteric virus primer EV1 to the above reverse transcription PCR reactants; performing the second PCR by adding Adenovirus primer AV3/AV4 and enteric virus primer EV3; and identifying the expanded virus DNA on the agarose gel. The both PCR consist of mutation step at 94deg.C for 4 minutes, expansion step with repeating the cycles of 30 second at 95deg.C, 30 second at 55deg.C and one minute at 72deg.C for 35 times, and final expansion step for 7 minutes at 72deg.C.