巢式PCR检测北京地区乙型肝炎病毒基因型的分布

Detection for Epidemic of Hepatitis B Virus Genotypes by Nested PCR with Multiplex Pairs Genotype-specific Primers

目的了解北京地区乙型肝炎病毒(HBV)感染者HBV基因型分布情况。方法从前S1基因到S基因中的保守序列设计出10条内外引物,将其中8条内引物分成A、B组,分别扩增A、B、C、D、E、F型HBV,根据PCR产物片断判定HBV基因型。结果用此种方法检测HBV感染的不同转归类型患者血清728份,对HBV DNA阳性标本进行基因型分析。HBV DNA阳性者276例,其中自限性感染者阳性率为5.8%(14/243),慢性无症状HBV携带者血清HBV阳性率为42.6%(113/265),慢性肝炎患者HBV阳性率为67.7%(149/220)。慢性肝炎组HBV DNA检出率与其他2组比较差异有统计学意义(P<0.05)。在276例HBV DNA阳性标本中,各基因型在不同组分布不同,但差异无统计学意义。HBV基因型总检出率以C型为主,占76.8%,B型占21·7%,同时检出B型和C型为1.5%,未检出A、D、E、F基因型。结论北京地区感染HBV基因型为B型和C型,以C型为主,感染结局不同组间基因型分布比较差异无统计学意义。

Objective To investigate the epidemic of HBV genotypes among HBV-infected persons in Beijing. Methods Ten outer and inner primers were designed on the basis of the conserved nature of nucleotide sequences in regions of the Pre-S1 through S genes, in which 8 inner primers were divided into mix A and B to amplify HBV OF genotype A, B,C and D, E, F respectively. Genotypes of HBV were determined directly by the size of PCR products. Results The HBV DNA in 728 sera samples collected from HBV-infected patients with different outcome were tested and genotyped by this nested PCR. 276 samples （37.9%） were HBV DNA positive. 5. 8% （14/243） self-limited HBV-infected patients were HBV-DNA positive, while chronic asymptomatic HBV carriers 42.6% （113/265）, chronic viral hepatitis 67.7% （149/220）respectively. There was significant difference between the chronic viral hepatitis and the other two groups. Among the 276 HBV DNA positive samples, the percentage of genotype C was 76.8%, type B 21.7 %, mixed genotype B and genotype C 1.5%, while none of genotype A, D, E and F had been found. Conclusion The HBV genotypes prevailed among HBV-infected persons in Beijing are genotype B and C, and the major genotype is genotype C. There is no statistical difference between the groups.