Detection and quantitation of infectious human adenoviruses and JC polyomaviruses in water by immunofluorescence assay |
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Authors: | Byron CalguaCelia Regina Monte Barardi Silvia Bofill-MasJesus Rodriguez-Manzano Rosina Girones |
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Institution: | a Department of Microbiology, Faculty of Biology, University of Barcelona, Diagonal 645, 08028 Barcelona, Spain b Department of Microbiology, Immunology and Parasitology, Center of Biological Sciences, Universidade Federal de Santa Catarina, 88040-900 Florianópolis, Santa Catarina, Brazil |
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Abstract: | Human adenoviruses (HAdV) and JC polyomaviruses (JCPyV) have been proposed as markers of fecal/urine contamination of human origin. An indirect immunofluorescence assay has been developed to quantify infectious human adenoviruses types 2 and 41 and JC polyomaviruses strain Mad-4 in water samples. The immunofluorescence assay was compared with other quantitative techniques used commonly such as plaque assay, tissue culture infectious dose-50 and quantitative PCR (qPCR). The immunofluorescence assays showed to be specific for the detection of infectious viruses, obtaining negative results when UV or heat-inactivated viruses were analyzed. The assays required less time and showed higher sensitivity for the detection of infectious viral particles than other cell culture techniques (1 log10 more) evaluated. River water samples spiked previously with human adenoviruses and raw sewage samples were also analyzed using the proposed immunofluorescence assay as well as by qPCR. The results show quantitations with 2 log10 reduction in the numbers of infectious viruses compared with the number of genome copies detected by qPCR. The immunofluorescence assay developed is fast, sensitive, specific, and a standardizable technique for the quantitation and detection of infectious viruses in water samples. |
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Keywords: | Immunofluorescence assay Plaque assay TCID50 qPCR Cell culture Adenoviruses JC polyomavirus |
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