The natural genomic variability of poliovirus analyzed by a restriction fragment length polymorphism assay

doi:10.1016/0042-6822(91)90434-D

Virology

Volume 184, Issue 2, October 1991, Pages 645-654

https://doi.org/10.1016/0042-6822(91)90434-D Get rights and content

Abstract

The genomic variability of poliovirus was examined by analyzing the restriction fragment length polymorphism of a reverse-transcribed genomic fragment amplified by the polymerase chain reaction. The fragment was a 480-nucleotide sequence of the poliovirus genome coding for the N-terminal half of the capsid protein VP1, including antigenic site 1. The identification of a pair of generic primers flanking this fragment allowed its amplification in practically all the poliovirus strains tested so far (more than 150). By using the restriction enzymes HaeIII, Ddel, and HpaII, strain-specific restriction profiles could be generated for the amplified genomic fragment of each of the six reference poliovirus strains tested: one representative wild poliovirus of each of the three serotypes (P1/Mahoney, P2/Lansing, and P3/Finland/23127/84) and the three Sabin vaccine strains. When 21 poliovirus field isolates previously identified as Sabin vaccine-related were tested, they showed restriction profiles identical to those of the originating homotypic Sabin virus, demonstrating the conservation of these profiles during virus replication in humans. These profiles could thus be used as markers for Sabin-derived genotypes. To compare the distribution of poliovirus genotypes in nature before and after the introduction of poliovirus vaccines, the restriction profiles of the amplified genomic fragment of a total of 72 strains of various geographic and temporal origins were determined. Strains isolated before the introduction of polio vaccines displayed a wide diversity of genotypes. In contrast, wild (Sabin unrelated) strains isolated after vaccine introduction, during a single epidemic in a particular geographic area, showed identical or very similar restriction profiles, indicating the circulation of predominant regional genotypes. Our results indicate that the assay we developed for the analysis of the restriction fragment length polymorphism of the poliovirus genome may be used to identify and characterize poliovirus genotypes circulating in nature.

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VP1 RT-PCR was performed by using SuperScript III one-step RT-PCR system with Platinum Taq DNA Polymerase (Thermo Fisher Scientific, USA). Primer pair UG1/UC11 (Balanant et al., 1991) was used to amplify the entire VP1 coding region of PV isolates, and primer pairs 486/488, 040/011, 008/013, 197/011, UF1/UR1 and 040/DR1 (Oberste et al., 2000, 2006; Tao et al., 2014) were used for the amplification of entire VP1 sequence of NPEV strains. Purified PCR products (~900 nt in length) were sequenced bi-directionally with the BigDye Terminator v3.1 Cycle Sequencing kit (Applied Biosystems) and analyzed on an ABI 3130 genetic analyzer (Applied Biosystems).
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The natural genomic variability of poliovirus analyzed by a restriction fragment length polymorphism assay

Abstract

Virology

Lancet

Adv. Virus Res.

Virology

Virology

Virology

J. Biol. Stand.

J. Mol. Biol.

Differentiation of types of poliomyelitis viruses. III. The grouping of fourteen strains into three basic immunological types

Am. J. Hyg.

Molecular detection and identification of enteroviruses using enzymatic amplification and nucleic acid hybridization

J. Clin. Microbiol.

Synthetic peptides from four separate regions of the poliovirus type 1 capsid protein VP1 induce neutralizing antibodies

Molecular pathogenesis of type 2 poliovirus in mice

Eur. J. Epidemiol.

Determination of type 1 poliovirus subtype classes with neutralizing monoclonal antibodies

Dev. Biol. Stand.

Natural variation of poliovirus neutralization epitopes

Infec. lmmun.

Neutralization epitope patterns on poliovirus strains isolated from paralytic cases

Dev. Biol. Stand.

WHO collaborative study on the use of monoclonal antibodies for the intratypic differentiation of poliovirus strains

Bull. WHO

Three-dimensional structure of poliovirus at 2.9 A resolution

Science

The nucleotide sequence of a type 3 poliovirus isolated during a recent outbreak of poliomyelitis in Finland

J. Gen. Virol.

Monoclonal antibodies of four different specimens for neutralization of type 1 polioviruses

Infect. Immunol.

Individual-specific ‘fingerprints’ of human DNA

Nature (London)