Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Patent
1994-12-21
1998-03-17
Marschel, Ardin H.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
4241301, 4241311, 4241841, 4241931, 4241991, 4242041, 4242051, 4242171, 424 931, 424 932, 424520, 435 5, 435 6, 435 71, 435 911, 435 912, 435 915, 4352351, 435236, 435237, 435239, 435948, 436501, 436543, 436547, 436 8, 436811, 436819, 536 231, 536 241, 536 243, 536 2431, 536 2432, 536 2433, 935 77, 935 78, C12Q 168
Patent
active
057285195
ABSTRACT:
The present invention provides a method of classifying an unclassified live poliovirus vaccine as having an acceptable or unacceptable level of neurovirulence, comprising, prior to vaccine administration, the steps of: a) selectively amplifying a region of an unclassified poliovirus vaccine genome containing a nucleotide position predictive for increased neurovirulence using selectively mismatched primers, whereby a restriction endonuclease site in the selectively amplified region is created by introducing a site-specific mutation into the amplified region; b) digesting an amount of the amplified region with a restriction endonuclease that specifically cleaves the amplified sequences in revertant viruses which contain a reversion at the nucleotide position predictive for increased neurovirulence; c) digesting an amount of the amplified region with a restriction endonuclease that specifically cleaves the amplified sequences in nonrevertant viruses which contain the nucleotide position predictive for increased neurovirulence; d) quantifying the percentage of revertant viruses in the unclassified vaccine; and e) comparing the percentage of revertant viruses in the unclassified vaccine to the percentage of revertant viruses in an accepted reference vaccine, and unclassified vaccine with a highter percentage of revertant viruses than in the reference vaccine being classified as unacceptable or an unclassified vaccine with an equal or lower percentage of revertant viruses than in the reference vaccine being classified as acceptable. Related methods and kits are also provided.
REFERENCES:
Chumakov et al., "Assessment of the Viral RNA Sequence Heterogeneity for Control of OPV Neurovirulence," Brown, F., and Lewis, B.P. (eds): Poliovirus Attenuation: Molecular Mechanisms and Practical Aspects, Dev. Biol. Stand. Basel, Karger, vol. 78, pp. 79-89 (1993).
Lu et al., "Quantitative Aspects of the Mutant Analysis by PCR and Restriction Enzyme Cleavage (MAPREC)," PCR Methods and Applications 3:176-180 (1993).
Ren et al., "Identification of Two Determinants that Attenuate Vaccine-Related Type 2 Poliovirus," Journal of Virology 65(3):1377-1382 (Mar., 1991).
Macadam et al., "The 5' Noncoding Regions of the Type 2 Poliovirus Vaccine Strain Contains Determinants of Attenuation and Temperature Sensitivity," Virology 181:451-458 (1991).
Christodoulou et al., "Mapping of Mutations Associated with Neurovirulence in Monkeys Infected with Sabin 1 Poliovirus Revertants Selected at High Temperature," Journal of Virology 64(10):4922-4929 (Oct., 1990).
Kawamura et al., "Determinants in the 5' Noncoding Region of Poliovirus Sabin 1 RNA that Influence the Attenuation Phenotype," Journal of Virology 63(3):1302-1309 (Mar., 1989).
Pollard et al., "Nucleotide Sequence of a Neurovirulent Variant of the Type 2 Oral Poliovirus Vaccine," Journal of Virology 63(11):4949-4951 (Nov., 1989).
Moss et al., "Mapping of Attenuating Sequences of an Avirulent Poliovirus Type 2 Strain," Journal of Virology 63(5):1884-1890 (May, 1989).
Westrop et a., "Genetic Basis of Attenuation of the Sabin Type 3 Oral Poliovirus Vaccine," Virology 63:1338-1344 (Mar., 1989).
J. W. Almond, "The Attenuation of Poliovirus Neurovirulence," Ann. Rev. Microbiol 41:153-80 (1987).
Evans et al., "Increased neurovirulence associated with a single nucleotide change in a noncoding region of the Sabin type 3 poliovaccine genome," Nature 314(11):548-550 (Apr., 1985).
Chumakov Konstantin M.
Levenbook Inessa S.
Norwood Laurie P.
Roninson Igor
Marschel Ardin H.
The United States of America as represented by the Department of
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