Molecular detection and identification of enteroviruses using enzymatic amplification and nucleic acid hybridization
| Autor: | Steven Tracy, Nora M. Chapman, C J Gauntt, U Fortmueller |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
Microbiology (medical)
Genotype viruses Molecular Sequence Data Dot blot Coxsackievirus Polymerase Chain Reaction law.invention Nucleic acid thermodynamics law Enterovirus Infections Humans Polymerase chain reaction Enterovirus Southern blot Base Sequence biology Hybridization probe Gene Amplification Nucleic Acid Hybridization virus diseases DNA biology.organism_classification Virology Molecular biology Agarose gel electrophoresis Primer (molecular biology) DNA Probes Research Article |
| Zdroj: | Journal of Clinical Microbiology. 28:843-850 |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.28.5.843-850.1990 |
| Popis: | Analysis of enteroviral genomes has revealed that the 5' nontranslated region is highly conserved, providing consensus sequences for the design of oligonucleotides which should anneal to most, if not all, human enteroviral RNAs. We designed and used a pair of such generic primers to enzymatically amplify cDNA from coxsackievirus group B types 1 through 6, poliovirus types 1 through 3, 4 coxsackievirus A types, and 29 echoviruses. The polymerase chain reaction (PCR) products generated with these enteroviral primers were analyzed by agarose gel electrophoresis, Southern blotting, or slot blot hybridization. A genotype-specific PCR was used to detect coxsackievirus B3, to the exclusion of other enteroviruses, by using a coxsackievirus B3 genome-specific primer pair that was derived from sequences coding for part of a capsid protein. A technique is demonstrated by which individual genotypes, for which no sequence information is known, can be identified by high-criterion hybridization analysis following amplification with generic enterovirus PCR primers. |
| Databáze: | OpenAIRE |
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