Evaluation of Secondary Concentration Methods for Poliovirus Detection in Wastewater

Jill C Falman,John Scott Meschke,Alexandra L Kossik,David S Boyle,Christine S Fagnant-Sperati

doi:10.1007/s12560-018-09364-y

Abstract

Effective surveillance of human enteric viruses is critical to estimate disease prevalence within a community and can be a vital supplement to clinical surveillance. This study sought to evaluate simple, effective, and inexpensive secondary concentration methods for use with ViroCap™ filter eluate for environmental surveillance of poliovirus. Wastewater was primary concentrated using cartridge ViroCap filters, seeded with poliovirus type 1 (PV1), and then concentrated using five secondary concentration methods (beef extract-Celite, ViroCap flat disc filter, InnovaPrep® Concentrating Pipette, polyethylene glycol [PEG]/sodium chloride [NaCl] precipitation, and skimmed-milk flocculation). PV1 was enumerated in secondary concentrates by plaque assay on BGMK cells. Of the five tested methods, PEG/NaCl precipitation and skimmed-milk flocculation resulted in the highest PV1 recoveries. Optimization of the skimmed-milk flocculation method resulted in a greater PV1 recovery (106 ± 24.8%) when compared to PEG/NaCl precipitation (59.5 ± 19.4%) (p = 0.004, t-test). The high PV1 recovery, short processing time, low reagent cost, no required refrigeration, and requirement for only standard laboratory equipment suggest that the skimmed-milk flocculation method would be a good candidate to be field-validated for secondary concentration of environmental ViroCap filter samples containing poliovirus.

Highlights

Effective surveillance of human enteric viruses is critical to estimate disease prevalence within a community, as enteric viruses are responsible for the majority of acute waterborne diseases worldwide (Wyn-Jones and Sellwood 2001; Fong and Lipp 2005; Maunula et al 2009; Sinclair et al 2009)
Environmental surveillance can be a vital supplement to clinical surveillance by testing wastewater and wastewater-impacted surface waters for the presence of viruses, thereby capturing information on a community’s health (Hovi et al 2012; World Health Organization [WHO] 2015a)
Stocks of poliovirus type 1 (PV1) vaccine strains were prepared by confluent lysis of buffalo green monkey kidney (BGMK) cell monolayers (Sobsey et al 1978)

Summary

Introduction

Effective surveillance of human enteric viruses is critical to estimate disease prevalence within a community, as enteric viruses are responsible for the majority of acute waterborne diseases worldwide (Wyn-Jones and Sellwood 2001; Fong and Lipp 2005; Maunula et al 2009; Sinclair et al 2009). Gastrointestinal tract of their host and are shed at high concentrations in fecal matter (typically between 105 and 1011 virions per g stool) for up to several months (Salim et al 1990; Blacklow and Greenberg 1991). They can be detected in wastewater treatment plant influents and effluents as well as surface waters in areas with poor sanitation (Petrinca et al 2009; Gibson et al 2011; Masachessi et al 2017).

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Conclusion