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Methods to Monitor Molecular Consistency of Oral Polio Vaccine

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Book cover Poliovirus

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

Abstract

Replication of viruses leads to emergence of mutations and their content in viral populations can increase by selection depending on growth conditions. Some of these mutations have deleterious effect on vaccine safety, such as neurovirulent reversions in the 5′-UTR of attenuated Sabin strains of poliovirus. Their content in vaccine batches must be tightly controlled during vaccine manufacture to ensure safety of the product. This chapter describes a quantitative molecular procedure called mutant analysis by PCR and restriction enzyme cleavage (MAPREC) that is used to monitor content of neurovirulent revertants in Oral Polio Vaccine (OPV). The method can be used for quantitative analysis of any other mutation in a viral population.

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References

  1. WHO (2012) Recommendations to assure the quality, safety and efficacy of live attenuated poliomyelitis vaccine (oral). WHO technical report series, vol 904. World Health Organization, Geneva, Switzerland. pp 1–87

    Google Scholar 

  2. Evans DM et al (1985) Increased neurovirulence associated with a single nucleotide change in a noncoding region of the Sabin type 3 poliovaccine genome. Nature 314(6011):548–550

    Article  CAS  PubMed  Google Scholar 

  3. Cann AJ et al (1984) Reversion to neurovirulence of the live-attenuated Sabin type 3 oral poliovirus vaccine. Nucleic Acids Res 12(20):7787–7792

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Macadam AJ et al (1991) The 5′ noncoding region of the type 2 poliovirus vaccine strain contains determinants of attenuation and temperature sensitivity. Virology 181(2):451–458

    Article  CAS  PubMed  Google Scholar 

  5. Christodoulou C et al (1990) Mapping of mutations associated with neurovirulence in monkeys infected with Sabin 1 poliovirus revertants selected at high temperature. J Virol 64(10):4922–4929

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Chumakov K et al (1992) RNA sequence variability in attenuated poliovirus. In: Brown F, Chanock R, Ginsberg H, Lerner R (eds) Vaccines 92. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp 331–335

    Google Scholar 

  7. Chumakov K et al (1993) Assessment of the viral RNA sequence heterogeneity for control of OPV neurovirulence. Dev Biol Standard 78:79–89, discussion 88–79

    CAS  Google Scholar 

  8. Punia P, Cane P, Teo CG, Saunders N (2004) Quantitation of hepatitis B lamivudine resistant mutants by real-time amplification refractory mutation system PCR. J Hepatol 40(6):986–992

    Article  CAS  PubMed  Google Scholar 

  9. Troy SB et al (2011) Use of a novel real-time PCR assay to detect oral polio vaccine shedding and reversion in stool and sewage samples after a Mexican national immunization day. J Clin Microbiol 49(5):1777–1783

    Article  PubMed  PubMed Central  Google Scholar 

  10. Laassri M, Bidzhieva B, Speicher J, Pletnev AG, Chumakov K (2011) Microarray hybridization for assessment of the genetic stability of chimeric West Nile/dengue 4 virus. J Med Virol 83(5):910–920

    Article  PubMed  PubMed Central  Google Scholar 

  11. Amexis G et al (2001) Quantitative mutant analysis of viral quasispecies by chip-based matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Proc Natl Acad Sci U S A 98(21):12097–12102

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Proudnikov D et al (2000) Analysis of mutations in oral poliovirus vaccine by hybridization with generic oligonucleotide microchips. Biologicals 28(2):57–66

    Article  CAS  PubMed  Google Scholar 

  13. Bidzhieva B, Laassri M, Chumakov K (2011) MAPREC assay for quantitation of mutants in a recombinant flavivirus vaccine strain using near-infrared fluorescent dyes. J Virol Methods 175(1):14–19

    Article  CAS  PubMed  Google Scholar 

  14. Chumakov KM (1995) Reverse transcriptase can inhibit PCR and stimulate primer-dimer formation. PCR Methods Appl 4:62–64

    Article  Google Scholar 

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Correspondence to Konstantin M. Chumakov .

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Chumakov, K.M. (2016). Methods to Monitor Molecular Consistency of Oral Polio Vaccine. 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_14

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  • DOI: https://doi.org/10.1007/978-1-4939-3292-4_14

  • 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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