乙肝病毒核心相关抗原检测试剂盒的研发及临床检测

Development and clinical detection of hepatitis B core-related antigen test kit

摘要:

目的 研发一种检测乙肝病毒核心相关抗原（hepatitis B core-related antigen，HBcrAg）的定量检测试剂盒并评价其性能。方法 通过研究原材料抗体、优化制备工艺和调整反应体系建立HBcrAg化学发光免疫检测方法，组装成试剂盒后利用重组HBcrAg和临床血清进行检测性能评估，数据处理使用IBM SPSS Statistics 20软件。结果 该试剂盒标准曲线为y=143.37x+344.91，R²=0.999 8；检测灵敏度为1.43 pg/mL；健康人参考区间 ≤ 25.595 pg/mL；检测208例慢性乙型肝炎患者血清，所有（18/18）乙型肝炎病毒（hepatitis B virus，HBV）DNA10⁸~10⁴ IU/mL血清HBcrAg检出值都在这个临界值以上；92.31%（24/26）HBV DNA10³~10² IU/mL血清HBcrAg检出阳性；44.51%（73/164）HBV DNA低于检出限血清HBcrAg检出阳性。结论 本试剂盒能较好地检测慢性乙型肝炎患者血清中的HBcrAg水平，在辅助慢性乙型肝炎患者治疗效果监测和正确判断治疗终点方面具有重要应用价值。

Abstract:

Objective To develop a quantitative test kit for determining hepatitis B core-related antigen（HBcrAg），and to evaluate its performance.Methods A chemiluminescent immunoassay for HBcrAg was established by studying the raw antibody，optimizing the preparation process，and adjusting the reaction system，and after the kit was assembled，recombinant HBcrAg and clinical serum were used to evaluate its detection performance. IBM SPSS Statistics 20 software was used for data processing.Results The standard curve of the kit was y=143.37x+344.91 （R²=0.999 8），with a detection sensitivity of 1.43 pg/mL and a reference interval of ≤25.595 pg/mL for healthy people. Serum samples were tested for 208 patients with chronic hepatitis B，and serum HBcrAg level was above this critical value for all 18 serum samples with an HBV DNA level of 10⁸-10⁴ IU/mL； for the 26 serum samples with an HBV DNA level of 10³-10² IU/mL，24（92.31%） tested positive for serum HBcrAg； for the 164 serum samples with HBV DNA below the limit of detection，73 （44.51%） tested positive for serum HBcrAg.Conclusion This kit can determine the serum level of HBcrAg in patients with chronic hepatitis B and thus has an important application value in monitoring the treatment outcome of patients with chronic hepatitis B and correctly judging the endpoint of treatment.