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Cited by 68Genomics and Phylogenetic StudiesViral Infections and Immunology ResearchVirology and Viral Diseases

Massively parallel sequencing for monitoring genetic consistency and quality control of live viral vaccines

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Neverov, Alexander, Chumakov, Konstantin·2010·Proceedings of the National Academy of Sciences
DOI 10.1073/pnas.1012537107PMC PMC2993378
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Abstract

Intrinsic genetic instability of RNA viruses may lead to the accumulation of revertants during manufacture of live viral vaccines, requiring rigorous quality control to ensure vaccine safety. Each lot of oral poliovirus vaccine (OPV) is tested for neurovirulence in animals and also for the presence of neurovirulent revertants. Mutant analysis by PCR and restriction enzyme cleavage (MAPREC) is used to measure the frequency of neurovirulent mutations at the 5′ untranslated region (UTR) of the viral genome that correlate with the level of neurovirulence determined by the monkey neurovirulence test. However, MAPREC can only monitor mutations at a few genomic loci and miss mutations at other sites that could adversely affect vaccine quality. Here we propose to use massively parallel sequencing (MPS) for sensitive detection and quantification of all mutations in the entire genome of attenuated viruses. Analysis of vaccine samples and reference preparations demonstrated a perfect agreement with MAPREC results. Quantitative MPS analysis of validated reference preparations tested by MAPREC produced identical results, suggesting that the method could take advantage of the existing reference materials and be used as a replacement for the MAPREC procedure in lot release of OPV. Patterns of mutations present at a low level in vaccine preparations were characteristic of seed viruses used for their manufacture and could be used for identification of individual batches. This approach may represent the ultimate tool for monitoring genetic consistency of live viral vaccines.

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  • In Sabin2, A481G is rapidly fixed upon replication in vaccinees.

    “In Sabin2 a transition mutation therein, A481G, is rapidly fixed upon replication in vaccinees (Neverov and Chumakov, 2010).”

    From Engineering the Live-Attenuated Polio Vaccine to Prevent Reversion to Virulence (2020) · cites this paper as Neverov and Chumakov, 2010

    The Abstract (page not specified) identifies 481-G as a Sabin 2 mutant and says Sabin strains can accumulate neurovirulent mutants while growing in vaccine recipients. It does not say that A481G is rapidly fixed in recipients. The Results’ reference to rapid accumulation concerns a different mutation, 2493-U, in cultured cells. (LLM, conf=0.85)

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Authors
Neverov, A., & Chumakov, K.
Affiliations
  • Center for Biologics Evaluation and Research (US)
  • National Institutes of Health (US)
Corresponding author
Chumakov, Konstantin
ORCID
Chumakov, Konstantin
Venue
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences
Published
2010-11
Volume
107
Issue
46
Pages
20063-20068
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
DOI
10.1073/pnas.1012537107
PMC
PMC2993378
OpenAlex
W1993080258
Cited by
68
References
3
Citations by year
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FWCI
1.96
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Phylogenetic Studies
  • Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Viral Infections and Immunology Research
  • Health Sciences › Medicine › Epidemiology › Virology and Viral Diseases
Keywords
oral polio vaccine, live attenuated vaccine, vaccine quality control, lot release, 5' untranslated region mutation, next-generation sequencing
MeSH terms
Animals, Chlorocebus aethiops, HeLa Cells, Humans, Mutation, Poliovirus Vaccine, Oral, Quality Control, Temperature, Vero Cells, Genome, Viral, Sequence Analysis, DNA, Poliovirus, High-Throughput Nucleotide Sequencing

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