NAT技术对献血人群HBV核酸单阳标本的检测结果分析

Analysis of detection results of HBV nucleic acid single positive samples in blood donors by NAT technique

目的 分析金华地区无偿献血者乙型肝炎病毒(HBV)核酸单阳标本检测结果,探究核酸扩增检测(NAT)技术在血液筛查中的应用价值。方法 收集2019年9月至2021年5月金华市中心血站初筛合格献血者血液标本102 641例,选择酶联免疫吸附试验(ELISA)检测乙肝表面抗原(HBsAg)(-)、NAT检测HBsAg(+)的标本,进行时间分辨免疫荧光法(TRFIA)定量检测乙肝两对半和实时荧光PCR技术定量检测HBV DNA。结果 纳入的102 641例血液标本中,经ELISA检测HBsAg(-)、丙型肝炎病毒(HCV)(-)、人类免疫缺陷病毒(HIV)(-)、梅毒螺旋体(TP)(-)标本共101 943例,经NAT检测HBV核酸单阳标本148例,占0.15%。148例HBV核酸单阳标本经乙肝两对半定量检测和HBV DNA鉴别检测,有12例(8.11%)检出HBsAg(+)[HBsAg≥0.05 IU/mL,滴度为(0.103±0.057)IU/mL],HBV DNA鉴别检测结果均呈反应性;136例(91.89%)检出HBsAg(-)(HBsAg<0.05 IU/mL),其中64例(43.24%)经HBV DNA鉴别检测结果呈反应性,72例(48.65%)经HBV DNA鉴别检测结果无反应性。148例HBV核酸单阳标本中,有135例(91.22%)检出乙肝核心抗体(HBcAb)(+)。根据血清学结果可将148例标本分为12种模式,HBV血清学模式5、10 HBV DNA鉴别检测非反应性标本与反应性标本的占比比较,差异具有统计学意义(P=0.003、0.007),其余HBV血清学模式HBV DNA鉴别检测非反应性标本与反应性标本的占比无明显差异(P>0.05)。HBV DNA鉴别检测出非反应性乙肝表面抗体(HBsAb)(+)标本33例(45.83%),高于反应性的21例(27.63%),差异具有统计学意义(χ2=5.286,P=0.021)。HBV DNA鉴别检测非反应性HBsAb(-)+HBcAb(+)标本33例(45.83%),低于反应性的52例(68.42%),差异具有统计学意义(χ2=7.716,P=0.005)。64例HBsAg(-)/HBV DNA(+)标本的HBV DNA滴度为(1.302±0.082)IU/mL。HBV DNA滴度<200 IU/mL的标本占95.31%(61/64),HBV DNA滴度<20 IU/mL的标本占48.44%(31/64)。结论 NAT技术有效降低了HBV窗口期感染和隐匿性乙型肝炎病毒感染(OBI)的漏检率。NAT技术联合ELISA血清学检测能有效降低HBV经血传播风险,保障血液安全。

Objective To analyze the detection results of hepatitis B virus (HBV) nucleic acid single positive samples in voluntary blood donors in Jinhua area, and to explore the application value of nucleic acid amplification testing (NAT) technology in blood screening. Methods A total of 102 641 blood samples from primary screening qualified blood donors in Jinhua Central Blood Station from September 2019 to May 2021 were collected. Samples of hepatitis B surface antigen (HBsAg) (-) detected by enzyme-linked immunosorbent assay (ELISA) and HBsAg (+) detected by NAT were selected for second liver two half-and -half by time-resolved fluoroimmunoassay (TRFIA) and quantitative detection of HBV DNA by real-time fluorescent PCR. Results Among the 102 641 blood samples included, 101 943 were HBsAg (-), hepatitis C virus (HCV) (-), human immunodeficiency virus (HIV) (-) and treponema pallidum (TP) (-) by ELISA, and 148 were HBV nucleic acid single positive by NAT, accounting for 0.15%. A total of 148 cases of HBV nucleic acid single positive samples by second liver two half-and-half of semi-quantitative detection and HBV DNA differential detection, HBsAg (+) was detected in 12 cases (8.11%) [HBsAg≥0.05 IU/mL, titer was (0.103±0.057) IU/mL], the results of HBV DNA differential detection were all reactive;HBsAg (-) (HBsAg<0.05 IU/mL) was detected in 136 cases (91.89%), of which 64 cases (43.24%) were reactive and 72 cases (48.65%) were non-reactive by HBV DNA differential detection. Among 148 cases of HBV nucleic acid single positive samples, hepatitis B core antibody (HBcAb) (+) was detected in 135 cases (91.22%). According to the serological results, 148 samples can be divided into 12 modes, there were significant differences in the proportions of HBV serological mode 5 and 10 HBV DNA differential detection between non-reactive samples and reactive samples (P=0.003, 0.007), and there were no significant differences in the proportions of the rest of the HBV serological model HBV DNA differential detection between non-reactive samples and reactive samples (P>0.05). There were 33 cases (45.83%) of non-reactive antibody to hepatitis surface untigen (HBsAb) (+) samples were detected by HBV DNA differential detection, which was higher than 21 cases (27.63%) of reactive samples, and the difference was statistically significant (χ^(2)=5.286, P=0.021). There were 33 cases (45.83%) of non-reactive HBsAb (-) + HBcAb (+) samples were detected by HBV DNA differential detection, which was lower than 52 cases (68.42%) of reactive samples, and the difference was statistically significant (χ2=7.716, P=0.005). The HBV DNA titer of 64 HBsAg (-)/HBV DNA (+) samples was (1.302±0.082) IU/mL. The samples with HBV DNA titer<200 IU/mL accounted for 95.31% (61/64), and the samples with HBV DNA titer<20 IU/mL accounted for 48.44% (31/64). Conclusion NAT technology effectively reduces the missed detection rate of HBV window phase infection and occult HBV infection (OBI). NAT technology combined with ELISA serological detection can effectively reduce the risk of HBV blood transmission and ensure blood safety.