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目的探讨福州地区无偿献血者人群隐匿性乙型肝炎病毒感染(OBI)状况及其HBV基因型分布特征与S区氨基酸突变的情况。方法应用血清学方法与核酸检测技术(NAT),对福建省福州市2011年11月-2013年3月的102 866(人)份无偿献血者标本做常规HBs Ag筛查及HBV DNA检测,排除HBV血清学标志物乙肝两对半检测结果阴性标本,确定HBs Ag-HBV DNA+为OBI标本;应用实时荧光定量PCR技术对OBI标本做HBV DNA检测,S区基因采用巢式PCR扩增和序列测定,使用MEGA5.0软件对HBV基因分型和对S区氨基酸做突变分析。结果2011年11月-2013年3月福州地区无偿献血者标本共筛查出75例HBs Ag-HBV DNA+[0.073%(75/102 866)],其中OBI率0.064%(66/102 866);66例OBI标本中,抗-HBc阳性比例为93.94%(62/66),HBV DNA检出率18.18%(12/66),HBV DNA为(13-302)IU/m L,仅1例标本的HBV DNA>200 IU/m L,15.15%(10/66)的OBI标本扩增出S区基因序列,其中B型7例、C型3例;突变分析发现在这10例中有7例HBV S区氨基酸发生突变,而其中又有6例的HBs Ag抗原决定簇基因及周边主要亲水区域(MHR)发生氨基酸突变。结论福州地区无偿献血者人群中存在一定的OBI感染率,其中抗-HBc阳性者比例占多数,OBI感染者的病毒载量低;HBV基因型主要以B型为主,HBV S区尤其是MHR的氨基酸突变可能是造成OBI发生的主要原因之一。
Objective To investigate the status of occult hepatitis B virus infection (OBI) and the distribution of HBV genotypes and the amino acid mutations in S region of the blood donors in Fuzhou area. Methods A total of 102 866 volunteer blood donors from Fuzhou, Fujian Province from November 2011 to March 2013 were routinely screened for HBsAg and HBV DNA by serological method and nucleic acid detection technique (NAT) HBs Ag-HBV DNA + OBI was determined by negative HBV serology marker HBV DNA test, HBV DNA was detected by real-time fluorescence quantitative polymerase chain reaction (PCR) and nested PCR amplification and sequencing , MEGA5.0 software was used to genotype HBV and make amino acid mutation analysis in S region. Results A total of 75 cases of HBs Ag-HBV DNA + [0.073% (75/102 866)] were screened out from non-compensation blood donors in Fuzhou area from November 2011 to March 2013, with an OBI rate of 0.064% (66/102 866); Among 66 OBI specimens, the positive rate of anti-HBc was 93.94% (62/66), the detection rate of HBV DNA was 18.18% (12/66), HBV DNA was (13-302) IU / mL, only 1 specimen HBV DNA> 200 IU / m L, 15.15% (10/66) of the OBI specimens amplified S region gene sequences, including 7 cases of type B, 3 cases of type C; mutation analysis found that in these 10 cases, 7 cases Amino acid mutations were found in the HBV S region, and 6 of the HBsAg epitopes and the surrounding major hydrophilic regions (MHRs) were mutated. Conclusion There is a certain prevalence of OBI in the population of unpaid blood donors in Fuzhou area, of which the proportion of positive anti-HBc is predominant and the infection of OBI is low. HBV genotypes are mainly type B, HBV S, especially MHR Amino acid mutations may be one of the major causes of OBI.