Molecular characterization of polio from environmental samples: ISSP, the Israeli sewage surveillance protocol

Lester M. Shulman, Yossi Manor, Musa Hindiyeh, Danit Sofer, Ella Mendelson

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

10 Scopus citations

Abstract

Polioviruses are enteric viruses that cause paralytic poliomyelitis in less than 0.5% of infections and are asymptomatic in >90% infections of naïve hosts. Environmental surveillance monitors polio in populations rather than in individuals. When this very low morbidity to infection ratio, drops drastically in highly vaccinated populations, environmental surveillance employing manual or automatic sampling coupled with molecular analysis carried out in well-equipped central laboratories becomes the surveillance method of choice since polioviruses are excreted by infected individuals regardless of whether or not the infection is symptomatic. This chapter describes a high throughput rapid turn-around time method for molecular characterization of polioviruses from sewage. It is presented in five modules: (1) Sewage collection and concentration of the viruses in the sewage; (2) Cell cultures for identification of virus in the concentrated sewage; (3) Nucleic acid extractions directly from sewage and from tissue cultures infected with aliquots of concentrated sewage; (4) Nucleic Acid Amplification for poliovirus serotype identification and intratypic differentiation (discriminating wild and vaccine derived polioviruses form vaccine strains); and (5) Molecular characterization of viral RNA by qRT-PCR, TR-PCR, and Sequence analysis. Monitoring silent or symptomatic transmission of vaccine-derived polioviruses or wild polioviruses is critical for the endgame of poliovirus eradication. We present methods for adapting standard kits and validating the changes for this purpose based on experience gained during the recent introduction and sustained transmission of a wild type 1 poliovirus in Israel in 2013 in a population with an initial IPV vaccine coverage >90%.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages55-107
Number of pages53
DOIs
StatePublished - 1 May 2016

Publication series

NameMethods in Molecular Biology
Volume1387
ISSN (Print)1064-3745

Keywords

  • Environmental surveillance
  • Intratypic differentiation
  • Oral polio vaccine (OPV)
  • Plaque assay
  • Poliovirus
  • RNA purification
  • RT-PCR
  • Sequence analysis
  • Tissue culture
  • Vaccine-derived poliovirus (VDPV)
  • Wild poliovirus
  • qRT-PCR

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