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 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 101 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.
Pallin
R
R
School of Health Sciences, The University, Sunderland, UK.
Wyn-Jones
A P
AP
Place
B M
BM
Lightfoot
N F
NF
D000327:Adsorption; D001419:Bacteria / Q000302:isolation & purification; D001435:Bacteriophages / Q000302:isolation & purification; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification*; D029822:Enterovirus B, Human / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis; D011998:Recreation; D012623:Seawater / Q000382:microbiology / Q000821:virology; D012680:Sensitivity and Specificity; D012822:Silicon Dioxide; D014871:Water Microbiology
D012367:RNA, Viral; D012822:Silicon Dioxide