Comparison of Concentration Methods for Quantitative Detection of Sewage-Associated Viral Markers in Environmental Waters

• DOI: 10.1128/aem.03851-14
• Publication Date: 2015-01-10T00:00:26Z
• Authors: W. Ahmed; V. J. Harwood; P. Gyawali; J. P. S. Sidhu; S. Toze
• Keywords: Ultrafiltration (renal)

ABSTRACT Pathogenic human viruses cause over half of gastroenteritis cases associated with recreational water use worldwide. They are relatively difficult to concentrate from environmental waters due typically low concentrations and their small size. Although rapid enumeration by quantitative PCR (qPCR) has the potential greatly improve quality analysis risk assessment, upstream steps capturing recovering sources along removing inhibitors extracted nucleic acids remain formidable barriers routine use. Here, we compared efficiency virus recovery for three methods concentrating two microbial source tracking (MST) viral markers adenoviruses (HAdVs) polyomaviruses (HPyVs) one liter tap river samples on HA membranes (90 mm in diameter). Samples were spiked raw sewage, adsorption was promoted acidification (method A) or addition MgCl 2 (methods B C). Viral acid directly A), eluted NaOH concentrated centrifugal ultrafiltration No inhibition qPCR observed processed method A, but occurred C. Recovery efficiencies HAdVs HPyVs ∼10-fold greater A (31 78%) than C (2.4 12%). Further revealed that majority not membrane, resulting poor recovery. The modification originally published include a larger diameter membrane extraction kit could accommodate resulted concentration good lack inhibitory compounds. frequently used strategy absorption added cations (Mg 2+ ) elution inefficient more prone inhibition, will result underestimation prevalence waters.