Analysis of Mutations in Oral Poliovirus Vaccine by Hybridization with Generic Oligonucleotide Microchips
- Resource Type
- Authors
- Andrei D. Mirzabekov; Dmitry Proudnikov; Jerome A. Donlon; Eugene Kirillov; Konstantin Chumakov; Gennady V. Rezapkin
- Source
- Biologicals. 28:57-66
- Subject
- Photomicrography
Quality Control
DNA, Complementary
DNA Mutational Analysis
Population
Mutant
Bioengineering
Gene mutation
Biology
Polymerase Chain Reaction
Applied Microbiology and Biotechnology
chemistry.chemical_compound
Complementary DNA
Point Mutation
education
Oligonucleotide Array Sequence Analysis
Pharmacology
education.field_of_study
Virulence
General Immunology and Microbiology
Oligonucleotide
Point mutation
Viral Vaccine
Reproducibility of Results
General Medicine
Virology
Molecular biology
Poliovirus
Microscopy, Fluorescence
chemistry
Poliovirus Vaccine, Oral
Mutation
RNA, Viral
Safety
Software
DNA
Biotechnology
- Language
- ISSN
- 1045-1056
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.