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Molecular Characterization of Polio from Environmental Samples: ISSP, The Israeli Sewage Surveillance Protocol

  • Protocol
Poliovirus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1387))

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 %.

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Abbreviations

AFP:

Acute flaccid paralysis

AR:

Analytic reagent grade chemicals

bp:

Base pairs

BSL-2:

Biological safety level two

CDC:

Centers for Disease Control and Prevention, Atlanta, GA, USA

CPE:

Cytopathic effect—morphological changes in cells resulting from infection

Ct:

Cycle threshold, the cycle at which specific signal from the probe is first detected above the threshold of detection in qRT-PCR

FBS:

Fetal bovine serum

GPEI:

Global Poliovirus Eradication Initiative

IPV:

Inactivated polio vaccine (trivalent contains all three Salk serotypes)

ITD:

Intratypic differentiation, e.g., determining whether the isolate is vaccine-like, VDPV, or wild

MOI:

Multiplicity of infection

OPV:

Live oral poliovirus (trivalent contains all three Sabin serotypes bivalent contains Sabin serotypes 1 and 3, monovalent contains either serotype 1, 2, or 3, exclusively)

PCR:

Polymerase chain reaction

PFU:

Plaque forming units

qRT-PCR:

Quantitative reverse transcription polymerase chain reaction

RT-PCR:

Reverse transcription polymerase chain reaction

SIA:

Supplementary Immunization Activities

TD:

Typic differentiation determining the serotype of the virus of interest

VDPV:

Vaccine-derived polio virus

VP1:

Viral capsid protein 1

VP2:

Viral capsid protein 2

VP3:

Viral capsid protein 3

VP4:

Viral capsid protein 4

WHO:

World Health Organization

5′ UTR:

5′ untranslated region of polio and non-polio enteroviruses

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Author information

Authors and Affiliations

  1. Environmental Virology Laboratory, Central Virology Laboratory, Public Health Services, Chaim Sheba Medical Center, Tel Hashomer, 52621, Israel

    Lester M. Shulman Ph.D.

  2. Department of Epidemiology and Preventive Medicine, School of Public Health, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel

    Lester M. Shulman Ph.D., Musa Hindiyeh Ph.D., D(ABMM), MT(ASCP) & Ella Mendelson Ph.D.

  3. National Center for Viruses in the Environment, Central Virology Laboratory, Public Health Services, Chaim Sheba Medical Center, Tel Hashomer, 52621, Israel

    Yossi Manor M.Sc.

  4. Molecular Assay Section, Central Virology Laboratory, Public Health Services, Chaim Sheba Medical Center, Tel Hashomer, 52621, Israel

    Musa Hindiyeh Ph.D., D(ABMM), MT(ASCP)

  5. Central Virology Laboratory, Public Health Services, Chaim Sheba Medical Center, Tel Hashomer, 52621, Israel

    Danit Sofer Ph.D. & Ella Mendelson Ph.D.

Authors
  1. Lester M. Shulman Ph.D.
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  2. Yossi Manor M.Sc.
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  3. Musa Hindiyeh Ph.D., D(ABMM), MT(ASCP)
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  4. Danit Sofer Ph.D.
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  5. Ella Mendelson Ph.D.
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Corresponding author

Correspondence to Lester M. Shulman Ph.D. .

Editor information

Editors and Affiliations

  1. Division of Virology, NIBSC, Hertfordshire, United Kingdom

    Javier Martín

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Shulman, L.M., Manor, Y., Hindiyeh, M., Sofer, D., Mendelson, E. (2016). Molecular Characterization of Polio from Environmental Samples: ISSP, The Israeli Sewage Surveillance Protocol. In: Martín, J. (eds) Poliovirus. Methods in Molecular Biology, vol 1387. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3292-4_5

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  • DOI: https://doi.org/10.1007/978-1-4939-3292-4_5

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3291-7

  • Online ISBN: 978-1-4939-3292-4

  • eBook Packages: Springer Protocols

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