Molecular quantitative analysis of human viruses in California stormwater

• Published in: Water research (Oxford) Vol. 41; no. 19; pp. 4287 - 4298
• Main Authors: Rajal, V.B., McSwain, B.S., Thompson, D.E., Leutenegger, C.M., Wuertz, S.
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01.11.2007; Elsevier Science
• Subjects: Adenoviridae; Adenovirus; Applied sciences; bacteriophages; Base Sequence; California; coliform bacteria; detection limit; DNA Primers; Enterovirus; equations; Exact sciences and technology; feces; genome; Hollow fiber ultrafiltration; mathematical models; microbial genetics; new methods; nucleic acids; Other industrial wastes. Sewage sludge; Pathogen detection; Pollution; Polymerase Chain Reaction; Public health; quantitative analysis; Quantitative PCR; Reference Standards; storm drains; stormwater; total suspended solids; ultrafiltration; vertebrate viruses; Viruses - classification; Viruses - genetics; Viruses - isolation & purification; Wastes; Water Microbiology; water quality; Water treatment and pollution; California; United States; North America; America; USA, California; Hollow fiber ultrafiltration; Pathogen detection; Enterovirus; Adenovirus; Quantitative PCR; Public health; Chemical analysis; Microbiology; Quantitative PCR,Hollow fiber ultrafiltration,Adenovirus,Enterovirus,Pathogen detection,Public health; Coliforms; Indicator; Polymerase chain reaction; Membrane separation; Health and environment; Ultrafiltration; Detection limit; Pathogenic; Health hazards; Storm water; Biological contamination; Quantitative analysis
• ISSN: 0043-1354; 1879-2448
• DOI: 10.1016/j.watres.2007.06.002

Many human pathogenic viruses are transmitted via the oral–fecal route and water is one possible vector, representing a risk for public health. Sixty-one large-volume water samples from storm drains in California were processed by a two-step hollow fiber ultrafiltration procedure followed by molecular analysis for human enterovirus and adenovirus types. Each sample was spiked with a surrogate, the benign bacteriophage PP7. Both surrogate and human viruses were quantified by newly designed TaqMan ® PCR assays. Equations were developed that account for the main variables in the procedure: recovery of the ultrafiltration, efficiency of nucleic acid extraction, and effect of inhibitors on the amplification of viral targets. Adenovirus 40/41 was detected in one sample at 230 genomes per liter, and no other adenovirus or enterovirus types were found. Samples that resulted in nondetects are reported together with the corresponding sample-specific limit of detection ( S LOD), a useful tool when estimating the public health risk associated with the contact or ingestion of water. Virus concentrations did not correlate with traditional viable indicator concentrations or any of the physicochemical parameters measured. In contrast, coliform concentrations were correlated with total suspended solids. To our knowledge, this is the first study where all factors known to influence limits of detection have been investigated and integrated into equations that are widely applicable to the quantification of viruses or other microbial targets by PCR.