血液筛查乙型肝炎病毒表面抗原ELISA灰区检测及判定策略优化探讨

doi:10.3969/j.issn.1671-2587.2026.02.016

临床输血与检验 ›› 2026, Vol. 28 ›› Issue (2): 252-258.DOI: 10.3969/j.issn.1671-2587.2026.02.016

血液筛查乙型肝炎病毒表面抗原ELISA灰区检测及判定策略优化探讨

蔡茵, 李建钧, 陈慧, 高智俊, 马丽敏, 蒋苓, 郑岚, 周国平, 王迅

上海市血液中心,上海 200051

收稿日期:2025-07-16 接受日期:2025-09-26 出版日期:2026-04-20 发布日期:2026-04-22
通讯作者: 王迅,主要从事感染性输血风险的预防控制等相关领域的研究,（E-mail）wangxun@sbc.org.cn。
作者简介:蔡茵,主要从事血液筛查检测相关研究,（E-mail）caiyin@sbc.org.cn。

Optimization and Decision Strategy for the ELISA Gray Zone in Hepatitis B Surface Antigen Screening in Blood Donation

CAI Yin, LI Jianjun, CHEN Hui, GAO Zhijun, MA Limin, JIANG ling, ZHENG Lan, ZHOU Guoping, WANG Xun

Shanghai Blood Center, Shanghai 200051

Received:2025-07-16 Accepted:2025-09-26 Online:2026-04-20 Published:2026-04-22

摘要/Abstract

摘要： 目的调查HBsAg酶联免疫吸附试验（ELISA）检测灰区样本的真阳性率,为全国采供血机构建立标准化的HBsAg ELISA灰区检测及判定策略提供参考依据。方法对近5年HBsAg ELISA检测灰区样本进行回顾性分析,并将近6个月的HBsAg ELISA检测灰区样本进行补充检测,包括额外2种HBsAg ELISA试剂检测,抗-HBc ELISA、抗-HBs ELISA检测及核酸定量检测,以及增加提取血浆量的核酸检测。结果比对4种HBsAg ELISA试剂检测灰区率,其中试剂3显著高于其他试剂,为1.56‰,不同实验室设备性能、不同试剂批号间也存在显著差异,同时发现献血者自身因素及标本性状与灰区结果也具有相关性。HBsAg灰区样本的核酸阳性率显著高于阴性样本的核酸阳性率,在HBsAg灰区且抗-HBc阳性的样本中核酸阳性率为10%,经病毒核酸定量检测,其病毒载量均远高于血液筛查核酸试剂中乙肝病毒核酸的最低检出限,另对7例常规核酸筛查为阴性HBsAg灰区抗-HBc阳性血浆袋样本,进行增加提取血浆量的核酸补充实验,均为阴性。结论采供血机构在使用灵敏度表现优秀的核酸检测系统的前提下可以取消HBsAg ELISA灰区设置,或者可以通过对HBsAg ELISA灰区样本增加抗-HBc检测,建立有效排除灰区样本中假反应性的检测策略,以保留珍贵的血液资源。

关键词: 献血者筛查, HBsAg检测, ELISA灰区, 抗-HBc检测, 核酸检测, 献血者保留

Abstract: Objective To investigate the true positive rate of hepatitis B surface antigen (HBsAg) enzyme-linked immunosorbent assay (ELISA) samples within the gray zone, and to propose a standardized strategy for processing such results in blood banks and transfusion services nationwide. Methods A retrospective analysis of HBsAg ELISA gray zone samples from the past 5 years was conducted. For samples from the past 6 months, supplementary testing was done using two additional HBsAg ELISA reagents, anti-HBc ELISA, anti-HBs ELISA, and quantitative nucleic acid testing. In cases with limited plasma volume, the extracted plasma volume for nucleic acid testing was increased. Results The gray zone rates of four HBsAg ELISA reagents were compared. Reagent 3 exhibited a significantly higher gray zone rate (1.56‰) than the others. Intermittent significant differences were noted in performance between different laboratory equipment and reagent batch numbers. Additionally, donor-specific factors and specimen characteristics were correlated with the gray area results. The positivity rate of nucleic acid in HBsAg gray zone samples was significantly higher than the total positivity rate of nucleic acid in routine ELISA-negative samples, with 10% of gray zone samples showing positive anti-HBc nucleic acid results. Quantitative viral nucleic acid testing revealed viral loads exceeding the minimum detection limit for hepatitis B virus in blood screening reagents. For 7 plasma bag samples with positive anti-HBc but negative results in routine nucleic acid screening, additional nucleic acid testing by increasing plasma volume showed all results to be negative. Conclusion Blood banks and transfusion services can eliminate the HBsAg ELISA gray zone on the premise of using nucleic acid detection systems with high sensitivity. Alternatively, incorporating anti-HBc testing for HBsAg gray zone samples may serve as an effective strategy to rule out false reactivity, thereby supporting the conservation of valuable blood resources.

Key words: Blood screening, HBsAg testing, ELISA gray zone, Anti-HBc testing, Nucleic acid testing, Blood donor reserve

中图分类号:

R446.61R512.6

蔡茵, 李建钧, 陈慧, 高智俊, 马丽敏, 蒋苓, 郑岚, 周国平, 王迅. 血液筛查乙型肝炎病毒表面抗原ELISA灰区检测及判定策略优化探讨[J]. 临床输血与检验, 2026, 28(2): 252-258.

CAI Yin, LI Jianjun, CHEN Hui, GAO Zhijun, MA Limin, JIANG ling, ZHENG Lan, ZHOU Guoping, WANG Xun. Optimization and Decision Strategy for the ELISA Gray Zone in Hepatitis B Surface Antigen Screening in Blood Donation[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2026, 28(2): 252-258.

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血液筛查乙型肝炎病毒表面抗原ELISA灰区检测及判定策略优化探讨

Optimization and Decision Strategy for the ELISA Gray Zone in Hepatitis B Surface Antigen Screening in Blood Donation

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