苏州地区抗-HBc阳性合格青年献血人群隐匿性HBV感染的血清学及分子生物学特性分析

目的探讨苏州地区乙型肝炎核心抗体(抗-HBc)阳性合格的青年献血人群隐匿性HBV感染的生物学及血清学特性。方法选取2013年10月至2016年6月乙型肝炎表面抗原(HBsAg)阴性、核酸检测(NAT)有反应性的青年献血者120例作为研究对象,进行乙型肝炎表面抗体(抗-HBs)定量检测和乙型肝炎两对半检测,对抗-HBc阳性标本进行病毒核酸提取和基本核心启动子区(BCP区)/前C区(PC区)及S区巢式PCR扩增,对扩增结果阳性标本进行基因测序及序列分析。结果 120例献血者中,抗-HBc(+)31例,其中22~25岁组25例,18~21岁组6例,差异有统计学意义(P0.05);抗-HBs(+)89例,其中22~25岁组16例,18~21岁组73例。利用巢式PCR扩增法对抗-HBc(+)标本进行PCR扩增,其中1例BCP区阳性,2例S区阳性,均属于22~25岁组。S区阳性标本的分型和测序结果显示:2例均为B型HBV,与野生型DNA序列相比,其中1例存在氨基酸序列E44U变异,1例存在T532G变异。结论对于HBsAg检测合格的抗-HBc(+)青年献血者,并不能保证其血液中一定不含有HBV DNA。为降低HBV经输血传播的风险,仍然需要进一步提高乙肝高流行地区核酸检测方法的灵敏度。

Analysis on serological and molecular biological characteristics of occult HBV infection among qualified young blood donors with anti-HBc positive in Suzhou area

Objective To explore the biological and serological characteristics of occult HBV infection among qualified young blood donors with anti-HBc positive in Suzhou area .Methods 120 young blood donors with negative HBsAg and reactivity in NAT in our hospital from October 2013 to June 2016 were selected as the research subjects .The anti-HBs quantitative detection and two pairs of semi-hepatitis B detection were performed .The samples of anti-HBc positive were conducted the viral nucleic acid extraction and nested PCR at the basic core promoter(BCP) region ,pre-core(PC) region and S region .The samples with positive amplification results were performed the gene sequencing and sequential analysis .Results Among 120 volunteer blood donors ,anti-HBc(+ ) had 31 cases ,in which 25 cases were in the 22-25 years old group and 6 cases in the 18-21 years old group ,the difference was statisti-cally significant(P<0 .05);anti-HBs(+ ) had 89 cases ,among them ,16 cases were in the 22 -25 years old group and 73 cases in the 18-21 years old group .The nest PCR was used to conduct PCR amplification in 31 samples of anti-HBc(+ ) ,in which 1 case was BCP region positive and 2 cases were S region positive ,all belonged to the 22-25 years old group .In the typing and sequencing for the samples of S region positive ,the results showed that all 2 cases were B type HBV .Compared with wild type DNA sequence , among them 1 cases had amino acid sequence E44U variation and 1 cases had T532G mutation .Conclusion For the anti-HBc(+ ) young blood donors qualified in HBsAg detection ,it is not guaranteed that their blood do not contain HBV DNA .It is still needed to further increase the sensitivity of nucleic acid detection method in hepatitis B highly prevalent areas fro decreasing the risk of HBV transmission by blood transfusion .