实时荧光PCR检测HBsAg阴性、抗HBc阳性献血者血液中HBV DNA研究

目的对乙型肝炎表面抗原（HBsAg）阴性、乙型肝炎核心抗体（抗HBc）阳性的献血者血液进行经输血传播乙型肝炎病毒（HBV）的风险评估，为完善HBV血液筛查模式及确保临床用血安全提供依据。方法对献血者血液常规检测阴性的标本进行8×45 μL汇集，应用实时荧光聚合酶链反应(PCR)进行混样核酸检测HBV DNA。对血液常规筛查和混样核酸检测合格的标本，进行随机乙型肝炎血清学5项标志物的酶联免疫吸附试验(ELISA)。对获得的HBsAg阴性、混样HBV DNA阴性、抗HBc阳性的标本，进一步采用单份样本（720 μL）实时荧光PCR法检测并定量分析。结果混样标本共检测12 552份，检出HBsAg阴性、HBV DNA阳性标本2份，阳性率为0.02%。随机筛查混样核酸检测阴性标本614份，检出抗HBc阳性标本320份，对此320份标本进行单份核酸检测，检出阳性标本1份，阳性率为0.31%。结论HBsAg阴性、抗HBc阳性献血者血液存在输血传播HBV的风险，应用核酸扩增技术检测血液HBV DNA能大大提高血液安全性。

Detection of blood HBV DNA in blood donors with HBsAg(-)/anti HBc(+) by real time fluorescence PCR

Objective To evaluate the risks of blood-transmitted HBV through blood donors with HBsAg（ - ）/an- ti-HBc（ ＋ ）, so as to improve better blood screening mode and ensure blood safety in clinical application. Methods Blood HBV DNA in serologically screened negative blood samples pooled at 8 - 45 μL size were detected by real-time fluorescence PCR, and PCR negative samples were tested for HBsAg, anti-HBs, HBeAg, anti-HBe and anti-HBc by ELISA. ,All HBsAg（ - ）, HBV DNA（ - ） and anti-HBc positive samples were detected by real-time fluorescence PCR individually for HBV DNA, HBV DNA positive samples were analysed by quantitative fluorescence PCR. Results Among 12 552 sero-negative samples, 2 were detected HBsAg（ - ）/HBV DNA（ ＋ ）, the positive rate was 0. 02%. Among 614 PCR negative samples, 320 were positive anti-HBc, and one of which was positive HBV DNA, the positive rate was 0. 31 %. Conclusion Donors with HBsAg（ - ）/anti-HBc（ ＋ ） remain potential risk for HBV transmission, and nucleic acid amplication technique can implement and improve blood safety.