摘要
目的探讨厦门地区乙型肝炎患者血清乙型肝炎病毒(HBV)基因型的分布情况。方法应用混合性型特异性引物聚合酶链反应法结合琼脂糖电泳,根据PCR产物片段大小直接判定HBV基因型;应用PCR扩增全S基因序列-DNA测序-生物信息学分析方法确定部分样本的亚基因型,并探讨前前S区编码的流行情况。结果在217例可以通过混合性型特异性引物聚合酶链反应法确定基因型的感染者中,B型83例(38.2%)、C型71例(32.7%),B/C混合型63例(29.0%)。HBeAg阳性患者中感染B基因型占50%,B/C型混合感染占22%;抗-HBe阳性患者中B/C型混合感染36%,B型38%;在慢性乙型肝炎患者中,以B基因型多见(46%),在肝硬化和肝癌患者中,以C(41%和42%)或B/C混合基因型(38%和33%)感染为主;20例患者血清经过DNA测序-生物信息学分析方法确定了亚基因型,其中B2亚型2例,C2亚型18例,其中混合性型特异性引物聚合酶链反应判断为B型的4例经过测序分析为C2亚型。B2亚型不编码前前S区,18例C2亚型均编码前前S区。结论本研究提示厦门地区乙型肝炎患者群的HBV基因型B、C型的感染率大致相同,亚基因型分析提示C2亚基因型占90%,可能是B/C基因型重组的结果。C2亚基因型病毒株均编码前前S区,提示前前S区的编码可能具有亚基因型特异性。
Objective To investigate the genotypes and subgenotypes of hepatitis B virus （HBV）in patients with HBV infection. Methods Mixed primers were designed to amplify genotype A,B,C or D,E,F of HBV,respectively. Electrophoresis of the second-round PCR products in a 3% agarose gel was employed to identify the genotypes. Furthermore,the whole S gene of HBV was cloned by PCR-TA clone method and the DNA sequence was analyzed by bioinformatics software to define subgenotype. Results Of the totally 217 patients with genotypes identified,83 （38.2%） were with genotype B,71（32.7%）genotype C and 63（29%）genotype B/C mixed infection; In patients with HbeAg positive, genotype B accounted for 50% and B/C mixed infection for 22%,while in patients with anti-Hbe positive,the B/C mixed infection was 36% and genotype B was 38%;In patients with chronic hepatitis B,the genotype B infection accounted for 46%,while in patients with ciiTohsis and liver cancer,the genotype C （41%,42%）and B/C infection （38%,33%）were common. In 20 isolates by bioinformatics software defined B2 subgenotype in 2 and C2 subgenotype in 18. All B/C mixed infection belonged to C2 subgenotype,and 4 isolates of C2 subgenotype were misjudged by multiple primers PCR method as genotype B. The pre-pre-S coding region existed in all 18 clones of C2 subtype,but none in B2 subtype. Conclusion The prevailed HBV genotypes among HBV-infected patients in Xiamen are B and C.
出处
《实用肝脏病杂志》
CAS
2008年第4期248-252,共5页
Journal of Practical Hepatology
基金
厦门市首批重大疾病科研攻关项目(WKZ0501)
厦门市卫生局医学科研立项项目(WSK0506)
厦门大学引进人才科研启动基金(Z03109)
福建省青年科技人才创新项目(2006F3127)
福建省高校新世纪人才创新资助项目
