Molecular detection and identification of enteroviruses using enzymatic amplification and nucleic acid hybridization

N. M. Chapman, S. Tracy, C. J. Gauntt, U. Fortmueller

Research output: Contribution to journalArticle

187 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)843-850
Number of pages8
JournalJournal of clinical microbiology
Volume28
Issue number5
StatePublished - May 8 1990

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Nucleic Acid Hybridization
Enterovirus
Human Enterovirus B
Polymerase Chain Reaction
Genotype
Genome
Poliovirus
Agar Gel Electrophoresis
Consensus Sequence
Capsid Proteins
Southern Blotting
Oligonucleotides
Complementary DNA
RNA

ASJC Scopus subject areas

  • Microbiology (medical)

Cite this

Molecular detection and identification of enteroviruses using enzymatic amplification and nucleic acid hybridization. / Chapman, N. M.; Tracy, S.; Gauntt, C. J.; Fortmueller, U.

In: Journal of clinical microbiology, Vol. 28, No. 5, 08.05.1990, p. 843-850.

Research output: Contribution to journalArticle

Chapman, N. M. ; Tracy, S. ; Gauntt, C. J. ; Fortmueller, U. / Molecular detection and identification of enteroviruses using enzymatic amplification and nucleic acid hybridization. In: Journal of clinical microbiology. 1990 ; Vol. 28, No. 5. pp. 843-850.
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