The Development of Molecular Detection Method and Differentiation of Genotypes of Enterovirus.
- Author:
Eun Soon KIM
;
Jung Hyun NAM
;
Yoo Kyum KIM
;
Ki Soon KIM
;
Jae Deuk YOON
- Publication Type:Original Article
- MeSH:
Central Nervous System Diseases;
Diagnosis;
Enterovirus B, Human;
Enterovirus Infections;
Enterovirus*;
Genotype*;
Humans;
Polymorphism, Restriction Fragment Length;
Polymorphism, Single-Stranded Conformational;
RNA
- From:Journal of the Korean Society of Virology
1997;27(2):169-176
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
In this study, the feasibility of identification and genotypic differentiation of enteroviruses was investigated by using nested reverse transcription-polymerase chain reaction (nested RT-PCR), single-stranded conformation polymorphism (SSCP), and restriction fragment length polymorphism (RFLP) techniques. Two hundred seventy-four clinical samples were assayed by both nested RT-PCR and tube culture method using MRC-5 and MK cells; 58 (86.6%) out of 67 enterovirus culture-positive samples contained enteroviral RNA. In addition, 114 (55.1%) of 207 samples from patients with suspected enteroviral CNS disease with negative viral cultures were positive by the nested RT-PCR. The nested RT-PCR products were genotyped by the SSCP method and the results were compared with serotypes. We could differentiate 6 subtypes, 3 of which are similar to coxsackievirus B3, B5, echovirus 11, plus 3 other subtypes. RFLP cleaved with Sty I, Bgl I, and Xmn I yielded characteristic patterns for each laboratory strains. This study demonstrates the usefulness of the RT-PCR for the rapid diagnosis of enterovirus infection and the potentials of the SSCP method for differentiation of enterovirus strains.
