The detection of enteroviruses in large volume concentrates of recreational waters by the polymerase chain reaction

A rapid and simple method was developed to detect enteroviruses in large-volume water samples. It relies on the adsorption of the virus capsids to silica particles under acidic conditions, allowing their recovery by relatively gentle centrifugation. Different reagents used in enterovirus concentrati...

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Bibliographic Details
Published in:Journal of virological methods 1997-08, Vol.67 (1), p.57-67
Main Authors: Pallin, R, Wyn-Jones, A.P, Place, B.M, Lightfoot, N.F
Format: Article
Language:English
Subjects:
Adsorption
Bacteria - isolation & purification
Bacteriophages - isolation & purification
Biological and medical sciences
coxsackievirus B5
Enterovirus
Enterovirus - genetics
Enterovirus - isolation & purification
Enterovirus B, Human - isolation & purification
Fundamental and applied biological sciences. Psychology
Hydrogen-Ion Concentration
Microbiology
PCR
Polymerase Chain Reaction - methods
Recreation
RNA, Viral - analysis
RT-PCR
Seawater - microbiology
Seawater - virology
Sensitivity and Specificity
Silicon Dioxide
Techniques used in virology
Virology
Water
Water Microbiology
Waterborne disease
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Summary:A rapid and simple method was developed to detect enteroviruses in large-volume water samples. It relies on the adsorption of the virus capsids to silica particles under acidic conditions, allowing their recovery by relatively gentle centrifugation. Different reagents used in enterovirus concentration and detection were seeded with Coxsackievirus B5 and used to optimise the recovery method, which was then used to detect the enteroviruses from seeded and unseeded 10 l seawater samples in one PCR tube rather than in up to 50 sub-sample volumes, demonstrating its use for routine environmental monitoring. Concentrates from 36 recreational water samples from three sites in N.E. England were analysed for enteroviruses by regular and the new method semi-nested PCR, and infectivity in cell culture. Some of the samples were also analysed for faecal indicator bacteria and F-specific bacteriophage. The results showed a marked increase in detection sensitivity when the whole sample concentrate was assayed as compared with a small volume aliquot.
ISSN:0166-0934
1879-0984
DOI:10.1016/S0166-0934(97)00076-1