Methods to Monitor Molecular Consistency of Oral Polio Vaccine

Methods Mol Biol. 2016:1387:263-77. doi: 10.1007/978-1-4939-3292-4_14.

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.

Keywords: Deep-sequencing; PCR; Quality control of Oral Polio Vaccine; Quantitative analysis of mutations; Reversion to virulence.

MeSH terms

  • Base Sequence
  • Genome, Viral
  • Humans
  • Mutation*
  • Poliomyelitis / prevention & control*
  • Poliomyelitis / virology*
  • Poliovirus / genetics*
  • Poliovirus / pathogenicity
  • Poliovirus Vaccine, Oral / adverse effects*
  • Poliovirus Vaccine, Oral / genetics*
  • Polymerase Chain Reaction / methods
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification
  • Restriction Mapping / methods

Substances

  • Poliovirus Vaccine, Oral
  • RNA, Viral