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Title: Analysis of mutations in oral poliovirus vaccine by hybridization with generic oligonucleotide microchips.

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.
Authors:
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Publication Date:
OSTI Identifier:
942596
Report Number(s):
ANL/BTC/JA-32828
Journal ID: ISSN 1045-1056; BILSEC; TRN: US200920%%60
DOE Contract Number:
DE-AC02-06CH11357
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biologicals; Journal Volume: 28; Journal Issue: 2000
Research Org:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org:
USDOD
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; CHARGE-COUPLED DEVICES; DETECTION; DNA; ELEMENTS; FLUORESCENCE; GENE MUTATIONS; HYBRIDIZATION; MIXTURES; MUTANTS; MUTATIONS; OLIGONUCLEOTIDES; QUALITY CONTROL; REVERTANTS; USES; VACCINES