Comparative study of three methods for genotyping hepatitis C virus strains in samples from Spanish patients

被引:48
作者
Forns, X
Maluenda, MD
LopezLabrador, X
Ampurdanes, S
Olmedo, E
Costa, J
Simmonds, P
SanchezTapias, JM
DeAnta, MTJ
Rodes, J
机构
[1] UNIV BARCELONA,HOSP CLIN & PROV,LIVER UNIT,E-08036 BARCELONA,CATALONIA,SPAIN
[2] UNIV EDINBURGH,SCH MED,DEPT MED MICROBIOL,EDINBURGH EH8 9AG,MIDLOTHIAN,SCOTLAND
[3] UNIV BARCELONA,HOSP CLIN & PROV,MICROBIOL LAB,E-08036 BARCELONA,CATALONIA,SPAIN
关键词
D O I
10.1128/JCM.34.10.2516-2521.1996
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Hepatitis C virus (HCV) genotypes may be investigated by a variety of laboratory methods that target different parts of the HCV genome and have various degrees of technical difficulty, Since the choice of a particular method is difficult, we compared the performance of (i) a type-specific PCR with type-specific primers from the core region, (ii) molecular hybridization of the PCR-amplified 5' noncoding region to type-specific probes, and (iii) identification of type-specific antibodies against epitopes of nonstructural region 4 by enzyme-linked immunosorbent assay (ELISA), One hundred fifty-one patients with biopsy-proved chronic hepatitis and HCV RNA in serum were investigated, The HCV genotype was identified in 99%, 100%, and 85% of the cases by type-specific PCR, probe hybridization, and ELISA, respectively. The type-specific PCR disclosed infection with type la in 3%, type Pb in 74%, and type 3a in 4% of the cases and suggested infection with two or more HCV types, including 2a/2c and 2b, in the remaining 18%. Apparently mixed infections were more prevalent in patients with past intravenous drug use (P < 0.001), but cloning and sequencing of PCR products did not confirm a mixed infection in any of the four cases investigated, Concordant results were obtained by the three procedures with virtually all of the samples in which the type-specific PCR revealed a single HCV genotype, Type-specific hy hybridization and ELISA usually recognized the genotype producing the strongest DNA band in samples in which type-specific PCR suggested a mixed infection, In conclusion, the three procedures evaluated in this study are reliable for investigation of HCV genotypes, Type-specific PCR provides information about HCV subtypes, but a mixed infection detected with this method should be interpreted with caution.
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页码:2516 / 2521
页数:6
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