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Evaluation of integrated cell culture‐PCR (C‐PCR) for virological analysis of environmental samples
Author(s) -
Greening G.E.,
Hewitt J.,
Lewis G.D.
Publication year - 2002
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.2002.01741.x
Subject(s) - infectivity , biology , polymerase chain reaction , enterovirus , real time polymerase chain reaction , reverse transcription polymerase chain reaction , virology , cell culture , microbiology and biotechnology , virus , messenger rna , gene , biochemistry , genetics
Aims: The aims of this study were to establish an integrated culture‐polymerase chain reaction (C‐PCR) method for detection of enteric viruses in environmental samples, and to evaluate it for sensitivity, speed and provision of virus infectivity data. Methods and Results : C‐PCR, direct reverse transcription (RT)‐PCR, PCR and plaque assay methods were used to detect enteroviruses and adenoviruses in seeded and naturally contaminated environmental samples. Using C‐PCR, infectious enterovirus presence was confirmed in 3 d and adenovirus presence in 5 d, compared with up to 10 d required by conventional cell culture methods. Conclusions: C‐PCR was the preferred method for detection of enteric viruses in environmental samples containing high viral concentrations. It was less successful for samples with low viral concentrations or containing toxic materials or inhibitors. Significance and Impact of the Study: C‐PCR provides sensitive, specific results within 2–5 d and is useful as a rapid screen for environmental samples of low toxicity.