MersV1, Main, Exploration, bibRecord, 003763

Analysis of Mutations in Oral Poliovirus Vaccine by Hybridization with Generic Oligonucleotide Microchips

Identifieur interne : 003763 ( Main/Exploration ); précédent : 003762; suivant : 003764

Analysis of Mutations in Oral Poliovirus Vaccine by Hybridization with Generic Oligonucleotide Microchips

Auteurs : Dmitry Proudnikov [États-Unis] ; Eugene Kirillov [Russie] ; Konstantin Chumakov [États-Unis] ; Jerome Donlon [États-Unis] ; Gennady Rezapkin [États-Unis] ; Andrei Mirzabekov [États-Unis]

Source :

Biologicals [ 1045-1056 ] ; 2000.

RBID : ISTEX:6E152F2F547A59ED7FC8C162DD47579EF1F3203A

English descriptors

Abstract

Abstract: This paper describes use of a new technology of hybridization with a micro-array of immobilized oligonucleotides for detection and quantification of neurovirulent mutants in Oral Poliovirus Vaccine (OPV). We used a micro-array consisting of three-dimensional gel-elements containing all possible hexamers (total of 4096 probes). Hybridization of fluorescently labelled viral cDNA samples with such microchips resulted in a pattern of spots that was registered and quantified by a computer-linked CCD camera, so that the sequence of the original cDNA could be deduced. The method could reliably identify single point mutations, since each of them affected fluorescence intensity of 12 micro-array elements. Micro-array hybridization of DNA mixtures with varying contents of point mutants demonstrated that the method can detect as little as 10% of revertants in a population of vaccine virus. This new technology should be useful for quality control of live viral vaccines, as well as for other applications requiring identification and quantification of point mutations.

Url:

https://api.istex.fr/ark:/67375/6H6-DQP3KRX7-P/fulltext.pdf

DOI: 10.1006/biol.1999.0241

Affiliations:

Russie, États-Unis

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Le document en format XML

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<front><div type="abstract" xml:lang="en">Abstract: This paper describes use of a new technology of hybridization with a micro-array of immobilized oligonucleotides for detection and quantification of neurovirulent mutants in Oral Poliovirus Vaccine (OPV). We used a micro-array consisting of three-dimensional gel-elements containing all possible hexamers (total of 4096 probes). Hybridization of fluorescently labelled viral cDNA samples with such microchips resulted in a pattern of spots that was registered and quantified by a computer-linked CCD camera, so that the sequence of the original cDNA could be deduced. The method could reliably identify single point mutations, since each of them affected fluorescence intensity of 12 micro-array elements. Micro-array hybridization of DNA mixtures with varying contents of point mutants demonstrated that the method can detect as little as 10% of revertants in a population of vaccine virus. This new technology should be useful for quality control of live viral vaccines, as well as for other applications requiring identification and quantification of point mutations.</div>
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Analysis of Mutations in Oral Poliovirus Vaccine by Hybridization with Generic Oligonucleotide Microchips

Analysis of Mutations in Oral Poliovirus Vaccine by Hybridization with Generic Oligonucleotide Microchips

Source :

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri