两种核酸血液筛查系统降低HBV残余风险的比较

发布时间：2018-03-05 17:52

本文选题：核酸检测　切入点：献血者筛查　出处：《中国输血杂志》2015年11期 　论文类型：期刊论文

【摘要】：目的评估不同原理核酸检测(NAT)血液筛查系统在降低输血传播HBV残余风险中的作用。方法对6 889人(次)无偿献血者血液标本做常规血清学检测,应用转录介导的扩增(TMA)技术(Ultrio试剂)对所有血样平行单样本定性检测HBV/HCV/HIV,对血清学无反应性标本5 165例采用PCR-荧光探针法(MPX试剂)以6人份混样模式做NAT。追踪2个NAT平台得到的反应性标本,罗氏CAP/CTM核酸定量检测系统做核酸鉴别试验,罗氏电化学发光做乙肝血清学5项检测,QPCR法测定HBV病毒载量,巢氏-PCR方法做HBV DNA基因分型,统计分析各项检测指标之间关联及差异。结果 TMA初筛检出HBV DNA单独反应性标本12例[反应率0.17%(12/6 889),鉴别试验阳性7例(占TMA初筛阳性数的58.3%)],PCR-荧光探针法拆分后检出HBV DNA单独反应性9例[0.17%(9/5 165)],2种NAT平台检测HBV DNA均为阳性4例,合计检出17例反应性标本。该2种原理的NAT检测系统对HBV DNA检测结果的一致性差(TMA vs QPCR,K0),检出率明显不同(TMA vs QPCR,P0.05)。阳性标本乙肝血清学5项检测:HBs Ag、HBe Ag均为阴性(0/16),抗-HBc阳性12例(12/16),抗-HBs阳性6例(6/16),抗-HBe阳性3例(3/16)。HBV DNA定量阳性4例,含量均5 IU/m L。献血者追踪结果:TMA检出HBV DNA单独反应性3例(3/10),鉴别试验HBV DNA阳性3例,MPX检出HBV DNA单独阳性5例(5/10),2种NAT平台检测均为阳性者3例,合计检出5例;10例追踪标本的HBs Ag、HBe Ag仍均为阴性,抗-HBc阳性8例(新转阳3例),抗-HBs阳性5例(新转阳1例),抗-HBe阳性2例(新转阳1例),HBV DNA定量阳性4例(新检出1例),含量均5 IU/m L。巢氏-PCR S区序列阳性标本做HBV基因分型:B型占66.7%(6/9),C型占33.3%(3/9)。结论 TMA与PCR-荧光定量法均能有效降低输血残余风险,但针对检出限附近低病毒载量标本均有部分漏检,血站在条件具备时可使用更高灵敏度的新试剂。
[Abstract]:Objective to evaluate the role of different principles of nucleic acid test (Nat) blood screening system in reducing the residual risk of transfusion transmitted HBV. Using transcription-mediated amplification technique (Ultrio reagent) to qualitatively detect HBV / HCV / HIV / HIV in parallel single sample of all blood samples and 5 165 cases of serological nonreactive specimens by PCR- fluorescence probe method (MPX). Reactive specimens obtained from the NAT platform, The CAP/CTM nucleic acid quantitative detection system was used to identify the nucleic acid, the serological detection of hepatitis B was performed by Roche electrochemiluminescence assay, and the carrying capacity of HBV virus was determined by QPCR. The genotyping of HBV DNA was performed by nested PCR method. Results 12 cases of HBV DNA single reactive specimen were detected by TMA. [the reaction rate was 0.17% and 12 / 6 889%, 7 cases (58.3%) were positive in differential test] PCR-fluorescence probe method was used to separate the sample. There were 9 cases of HBV DNA reactivity alone [0.17% / 5 165] and 4 cases were positive for HBV DNA detected by two kinds of NAT platforms. A total of 17 reactive specimens were detected. The results of the two principle NAT detection systems were not consistent with those of HBV DNA. The detection rate was significantly different from that of TMA vs QPCRX P0.05.The positive samples were positive for HBV serological tests, and all of them were 0 / 16% negative and anti-HBs Ag-HBeAg were negative. 12 cases were positive for -HBc, 6 cases were positive for anti-HBs, and 3 cases were positive for anti-HBe, 3 cases were positive for anti-HBe, 4 cases were positive for HBV DNA. The results of blood donor tracing showed that HBV DNA was only reactive in 3 cases (3 / 10), and HBV DNA was positive in 3 cases (P < 0 05). HBV DNA was positive in 5 cases (5 / 10) and HBV DNA in 3 cases (P < 0 05). In total, 5 cases (10 cases) were found to be still negative for HBs Ag-HBe Ag, and 10 cases were found to be still negative for HBe Ag. 8 cases were positive for anti-HBc (3 cases were newly converted to positive, 5 cases were positive for anti-HBs), 2 cases were positive for anti-HBe (1 case was newly converted to positive), 4 cases were positive for DNA (1 case was newly detected, all of them were positive in 5 IU/m / L). The proportion of HBV genotypes B to B was 66.7%. Conclusion the residual risk of blood transfusion can be effectively reduced by TMA and PCR- fluorescence quantitative method. However, some of the specimens with low viral load near the detection limit are missing, so the new reagent with higher sensitivity can be used in the blood station when the conditions are available.
【作者单位】：深圳市血液中心;
【基金】：深圳市科技创新委员会项目(JCYJ20140403093211510) 深圳市科卫生计生系统科研项目(201401074)
【分类号】：R512.62

【相似文献】