HBV DNA PCR检测在HBsAg阴性献血人群中的应用

Detection of HBV DNA by PCR on HBsAg negative blood donors

目的探讨HBsAg阴性献血者HBVDNA检测的应用价值，评估核酸检测的必要性。方法采用PCR检测HBsAg阴性献血者HBVDNA。采用8人份混合血样测定，超离心浓缩病毒，磁珠法提取病毒核酸。如HBVDNA为阳性，则进一步检测乙型肝炎病毒血清标志物5项。结果HBVDNA检测限量为25U／ml，23225份标本中有4份为HBVDNA阳性，检出率为0．17‰。进一步检测其他HBV感染的血清学指标，发现这4份标本中有2份为抗HBe和抗HBc阳性，1份为抗HBc阳性，1份为抗HBs、抗HBc阳性。对HBVDNA的定量测定表明，其含量在50～200U／ml。结论现行的2次酶联免疫技术的血液筛查存在HBV漏检，有必要在现有的血液筛查模式中增加抗HBc检测，或增加病毒核酸筛查。

Objective To define the application value of HBV DNA detection on HBsAg-negative blood donors and assess the necessity for nucleic acid detection. Methods Real-time PCR was used to detect HBV DNA on HBsAg negative blood donors. Pools of eight donor samples were used for NAT testing. Viruses were concentrated by centrifugation and the viral DNA extraction was performed using magnetic beads. If HBV DNA was positive, serological indicators including HBsAg, anti-HBs, HBeAg, anti-HBe, total anti-HBc was further detected. Results The HBV DNA detection limit was 25 U/ml. There were four HBV DNA positive cases among 23 225 specimens, and the detection rate was 0. 17‰. The further serological examination showed anti-HBe （ ＋ ）, anti-HBc （ ＋ ） in the two cases and anti-HBc （ ＋ ） in one case and anti-Hbs （ ＋ ）, anti-HBc （ ＋ ） in 1 case. The viral load can range form 50 to 200 U/ml. Conclusions The results indicate that there is false negative possibility in blood screening by ELISA. It is necessary to employ anti-HBc screening or NAT to blood donors screening.