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

AUTOR(ES)

Chapman, N M

RESUMO

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.

Documentos Relacionados

• Detection of enteroviruses by spot hybridization.
• Detection and identification of mycobacteria by DNA amplification and oligonucleotide-specific capture plate hybridization.
• Multicenter Proficiency Testing of Nucleic Acid Amplification Methods for the Detection of Enteroviruses
• Comparison of nucleic acid hybridization and nucleic acid amplification using conserved sequences from the 5' noncoding region for detection of bovine viral diarrhea virus.
• Detection of enteroviruses and rhinoviruses in clinical specimens by PCR and liquid-phase hybridization.