AWWA WQTC56977

AWWA WQTC56977 Detection of Enteric Viruses in Archived ICR Sample Concentrates Using an Integrated Cell Culture-Nested PCR Technique
Conference Proceeding by American Water Works Association, 11/01/2002

Warden, Paul S.; Ballester, Nicola A.; Morre, Amy E.; Fontaine, Justin H.; Fallon, Kristen S.; Margolin, Aaron B.

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The preamble of the Information Collection Rule (ICR) referenced the goal of archiving virus samples for future study. From samples submitted to our laboratory during the ICR, Analytical Services, Inc. (ASI) and University of New Hampshire (UNH) created a repository of sample concentrates that were representative of a broad geographic area and were consistent with respect to methods of sample collection, handling and storage. The objective of this study was to examine viral occurrence in archived source water sample concentrates using a sophisticated, sensitive molecular technique and to compare these results to the respective data obtained from the Total Cultivatable Virus Assay (TCVA) used in the ICR. Sample concentrates were analyzed using Integrated Cell Culture/nested Polymerase Chain Reaction (ICC/nPCR) for the detection of adenovirus 40 and 41, astrovirus, rotavirus and enteroviruses (coxsackie virus, echovirus and poliovirus). Of the 91 samples selected for inclusion in the study, 41 were positive by the TCVA method. In contrast, 50 of 91 samples (an increase of 22%) were positive for one or more viral types using ICC/nPCR. Of these 50 ICC/nPCR positive samples, 34 were confirmed to contain one or more types of infectious viruses. Significantly, over half of the TCVA-negative samples (29/50 or 58.0%) were positive for one or more viral types by ICC/nPCR. Sixteen of these samples (16/50 or 32.0%) were confirmed by ICC/nPCR to contained infectious viruses. Interestingly, of the 41 TCVA-positive samples, only 21 (51.2%) were positive by ICC/nPCR; the other 20 were negative. This could be a result of the age of the sample (samples had been stored at -80o C for 2-4 years); primer selection (reovirus primers were not included yet reoviruses are supported on BGMK cells); and/or the presence of other non-human viruses known to cause CPE on BGMK cells (not detected by the primers used in our assay). No rotavirus amplicons were detected in any samples, which may indicate their absence, their lack of stability in long term storage and/or during sample thawing and processing. Includes 5 references, tables, figures.