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目的了解安徽、福建、江西三地血液中心无偿献血者隐匿性乙型肝炎病毒(OBI)感染情况,探讨HBV基因分型特征及S区氨基酸变异特点。方法对2013年1月-2014年5月安徽省血液中心、2013年6月-2013年11月、2014年3月-2014年12月福建省血液中心、2013年6月-2013年9月江西省血液中心乙肝表面抗原酶联免疫法检测阴性(HBs Ag-)、核酸HBV鉴别试验阳性(NAT+)的102例无偿献血者标本进行抗-HBc检测,同时应用巢式PCR方法对这些标本进行HBV S区扩增并测序,采用MEGA 6软件进行HBV基因分型及S区氨基酸突变分析。结果在安徽省血液中心123 046例无偿献血者中筛查出21例HBs Ag-NAT+无偿献血者标本,OBI感染率为0.017%(21/123 046),其中76.2%(16/21)为抗-HBc阳性,这21例标本中,15例扩增出HBV S区片段,8例B型,7例C型;福建省血液中心HBs Ag-NAT+无偿献血者标本共51例,76.5%(39/51)为抗-HBc阳性,其中16例扩增出HBV S区片段,14例B型,2例C型;江西省血液中心HBs Ag-NAT+献血者标本共30例,80%(24/30)为抗-HBc阳性,其中4例扩增出HBV S区片段,1例B型,3例C型。35例扩增产物的OBI标本中,74.3%(26/35)出现HBV S基因MHR区氨基酸置换突变,主要集中在乙型肝炎病毒“α”决定簇区域。结论安徽地区无偿献血者OBI感染率为0.017%,福建及江西地区也有一定OBI感染;成功扩增HBV S区的OBI标本中,抗-HBc阳性率为安徽73.3%、福建93.8%、江西100%;B型为主要HBV基因型;HBV S基因MHR区变异,尤其是HBs Ag“α”决定簇区置换突变,可能是OBI形成原因之一。
Objective To investigate the prevalence of occult hepatitis B virus (OBI) in blood donors in blood centers in Anhui, Fujian and Jiangxi provinces and to explore the characteristics of HBV genotyping and amino acid variation in S region. Methods From January 2013 to May 2014, Anhui Provincial Blood Center, June 2013-November 2013, March 2014-December 2014 Fujian Provincial Blood Center, June 2013-September 2013 Jiangxi The detection of anti-HBc in 102 blood donors with HBsAg-positive and NAT-positive HBV antibodies was detected by the Hepatitis B virus surface antigen ELISA at the provincial center of blood and the HBV DNA was detected by nested PCR S region was amplified and sequenced. MEGA 6 software was used to analyze HBV genotype and S region amino acid mutation. Results Totally 21 HBs Ag-NAT + blood donors were collected from 123 046 blood donors in Anhui Blood Center. The infection rate of OBI was 0.017% (21/123 046), of which 76.2% (16/21) were anti-HBs -HBc was positive. Of the 21 specimens, 15 were positive for HBV S region, 8 were type B, and 7 were type C; 51 were HBsAg-NAT + blood donors in Fujian Blood Center, 76.5% / 51) were positive for anti-HBc, including 16 cases of HBV S region, 14 cases of type B and 2 cases of type C; 30 cases of HBsAg-NAT + blood donors in Jiangxi Blood Center, 80% 30) were anti-HBc positive, of which 4 cases amplified HBV S region fragment, 1 case of B type, 3 cases of C type. Of the 35 OBI samples, 74.3% (26/35) showed amino acid substitutions in the MHR region of HBV S gene, mainly in the region of the “α” determinant of hepatitis B virus. Conclusion The OBI infection rate of unpaid blood donors in Anhui Province was 0.017%, and OBI infection was also found in Fujian and Jiangxi Provinces. The positive rate of anti-HBc in OBI specimens successfully amplified in HBV S region was 73.3% in Anhui, 93.8% in Fujian and 100% ; Type B is the main HBV genotype; HBV S gene MHR region variation, especially HBsAg “α” determinant region substitution mutation may be one of the reasons OBI formation.