Genotyping of hepatitis E virus by PCR combining with single restriction endonuclease analysis / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 179-181, 2005.
Artigo
em Chinês
| WPRIM
| ID: wpr-333068
ABSTRACT
<p><b>OBJECTIVE</b>To develop a simple method for genotyping of hepatitis E virus (HEV) and to investigate HEV genotype distribution in Nanjing area.</p><p><b>METHODS</b>Twenty-seven full HEV sequences currently-available in GenBank were analyzed with MegAlign and MapDraw programs of DNA STAR software. Degenerate primers were designed and applied to amplify a fragment in HEV ORF1 region. HEV genotypes were determined by the size of the PCR products and by single restriction endonuclease analysis.</p><p><b>RESULTS</b>The PCR products of HEV genotype 1 and 2 were 275 bp and 269 bp in size. Distinctively, the PCR products of genotype 3 and 4 were 317 bp and 314 bp in size. Moreover, the PCR products of genotype 1 could be digested by Nae 1, but the products of genotype 2 could not. Distinctively, the PCR products of HEV genotype 3 could be digested by Not 1, but the products of genotype 4 could not. Six HEV reference strains standing for different HEV genotypes were clustered into their own types as predicted. Within 43 HEV IgM-positive clinical specimens collected in Nanjing, 19 were HEV PCR-positive and identified as genotype 4.</p><p><b>CONCLUSION</b>A simple method of PCR combined with single restriction endonuclease analysis is developed for HEV genotyping. This assay allows rapid identification of a large number of HEV isolates directly from clinical specimens. Among patients with hepatitis E in Nanjing, most were infected with HEV genotype 4.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Sangue
/
RNA Viral
/
Desoxirribonucleases de Sítio Específico do Tipo II
/
Enzimas de Restrição do DNA
/
Reação em Cadeia da Polimerase
/
Vírus da Hepatite E
/
Hepatite E
/
DNA Complementar
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Alergia e Imunologia
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Experimental and Clinical Virology
Ano de publicação:
2005
Tipo de documento:
Artigo
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