【摘要】 目的应用型特异性引物聚合酶链反应法(PCR)进行乙型肝炎病毒基因分型并分析该法的可靠性。方法应用型特异性引物PCR和INNO-LiPA分别对深圳、长春、北京152份HBV DNA阳性慢性乙型肝炎患者的血清标本进行了基因型分型,对该两种分型法不一致的血清标本再进行S区基因测序分型,以确定该两法的可靠性。结果型特异性引物PCR和INNO-LiPA的总符合率为86．8％(132／152),不一致率为13．2％(20／152)。型特异性引物PCR检测到81份(53．3％)B型;58份(38．2％)C型;13份(8．5％)B+C型混合感染,未检出其他基因型或混合感染的基因型。INNO-LiPA检测到74份(48．7％)B型;61份(40．1％)C型;5份(3．3％)B+C型混合感染;另检出3份(2．0％)A +B型混合感染;1份(0．7％)B+E型混合感染;1份(0．7％)C型与D型,1份(0．7％)D型感染,3份(2．0％)B／C／D型及3份未能分型。20份两法分型不一致的标本中,6份无剩余血清,对其余14份进行了S区基因测序分型,结果型特异性引物PCR与S区基因测序分型法的符合率为71．4％(10/14),而INNO-LiPA与S区测序法的符合率仅为7．1％(1／14),前者明显高于后者(P<0．05)。结论型特异性引物PCR和INNO-LiPA均可鉴定HBV基因型,但前者较为简便和可靠,且费用较低,可用于临床标本的检测和流行病学调查。更多还原

【Abstract】 Objective To determine the accuracy of a type-specific primer polymerase chain reaction (PCR) for identifying HBV genotypes. Methods A type-specific primer PCR and INNO-LiPA assay were used to identify HBV genotype of 152 serum samples of patients with chronic hepatitis B from Shenzhen, Changchun and Beijing. Fourteen of 20 serum samples with discrepant genotype were analyzed by the sequencing of HBV S gene to determine their accuracy. Results The total coincidence rate between the type-specific primer PCR and INNO-LiPA was 86.8%(132/152), with the discrepancy of 13.2%(20/152). Of 152 samples from patients with HBV infection determined by the type-specific PCR, the proportion of genotype B, C, and B+C were 53.3%(81/152), 38.2%(58/152) and 8.5%(13/152), respectively. The figures for INNO-LiPA were 48.7%(74/152), 40.1% (61/152) and 3.3% (5/152), respectively. In addition, 3(2.0%) genotype A + B, 1(0.7%) B + E, 1 (0.7%) C+D, 1(0.7%) D, 3(2.0%) B+C + D, and 3(2.0%) were unidentified by INNO-LiPA. Of 20 sera with discrepant genotype results by the type-specific primer PCR and INNO-LiPA, 6 had no serum left, and the remaining 14 samples were further analyzed by the sequencing of HBV S gene for genotyping. The total coincidence rate between the type-specific primer PCR and the sequencing of HBV S gene was 71.4% (10/14), which was significantly higher than that of INNO-LiPA(7.1 % , 1/14), P < 0.05. Conclusion Type-specific primer PCR and INNO-LiPA can be used for HBV genotyping. However, the type-specific PCR is more simple and accurate, with low cost. It may be used for test of clinical specimens and for epidemiologiesl investigations.更多还原