厦门市乙型肝炎患者病毒基因型的分析

Preliminary study on hepatitis B virus genotypes in Xiamen city

目的:采用多对型特异性引物,通过巢式PCR法检测厦门市乙型肝炎患者血清中乙型肝炎病毒(HBV)基因型的分布情况.方法:收集250例HBV感染患者血清,提取血清中HBV DNA作为模板,设计HBV前S1基因和S基因中区域内设计出10条内外引物,并将其中8条型特异性内引物分成A,B两组,分别扩增A,B,C和D,E,F型HBV,然后将第2轮PCR产物以用30g/L琼脂糖进行电泳,根据PCR产物电泳显示的产物长度判定HBV基因型,以了解厦门HBV基因型分布情况.结果:共120例确定了HBV基因型.患者群中慢性乙型肝炎90例,占75.0%,急性乙型肝炎、肝炎肝硬化、原发性肝癌分别占5.8%(7/120)、6.7%(8/120)和12.5%(15/120).分型结果:B型58例(48.3%)、C型30例(25.0%),B/C混合型32例(26.7%).HBeAg阳性患者中B基因型占63.8%,B/C型混合感染21.9%;抗-HBe阳性患者中以B/C型混合感染68.8%,B型25.9%,HBeAg阳性组与抗-HBe组之间比较发现B型和B/C混合型之间(P<0.05).结论:厦门乙型肝炎患者HBV基因型以B型为主,B/C混合感染是一个值得重视的问题.

AIM： To investigate the genotypes of hepatitis B virus （HBV） in Xiamen city by nested polymerase chain reaction （PCR） with multiplex pairs of genotype-specific primers. METHODS： A total of 250 HBV-infected patients were included in this study. The serum samples were collected and the serum HBV DNA was used as templates. Ten outer and inner primers were designed on the basis of nucleotide sequences in the regions of Pre-S1 and S genes, of which 8 genotype-specific inner primers were divided into 2 groups： A and B. Genotype A, B, C or D, E, F of HBV were amplified, respectively. The genotypes of the second-round PCR products were identified using agarose gel electrophoresis （30 g/L）. RESULTS： Of the 250 selected patients, 120 received the above genotyping successfully. There were 90 （75.0%） diagnosed with chronic hepatitis B, 7 （5.8%） with acute hepatitis B, 8 （6.7%） with liver cirrhosis, and 15 （12.5%） with hepatocellular carcinoma. Of the 120 cases, 58 （48.3%） were found with genotype B, 30 （25.0%） with genotype C and 32 （26.7%） with genotype B/C. In the patients with positive HBeAg, genotype B and B/C were confirmed in 63.8% and 21.9% of them, respectively. However, the patients with genotype B/C and B accounted for 68.8% and 25.9% of the anti-HBe-positive patients. There were significant differences between the 2 subgroups in the genotype distribution （P 〈 0.05）. CONCLUSION： Genotype B prevails among HBV-infected patients in Xiamen city, and type B/C HBV infection a considerable problem.