精子细胞乙型肝炎病毒核心抗原流式细胞检测方法的建立

Establishment of flow cytometry detection of HBcAg in sperm cells from hepatitis B patients

目的探讨建立流式细胞分析术对精子细胞乙型肝炎病毒核心抗原(HBcAg)定性、定量检测的诊断方法。方法收集62例乙型肝炎患者精液标本,制备成精子细胞单细胞悬液,用荧光标记的小鼠抗HBcAg单克隆抗体与标本细胞充分反应后,应用流式细胞分析术进行定性、定量检测,并制定诊断标准。结果该方法对乙型肝炎患者精子细胞HBcAg定性阳性检出率即敏感度为96.77%,对10例健康对照标本HBcAg定性阴性检出率即特异度为100%,阳性、阴性预测值均为100%,准确度97.22%。该方法精子细胞中HBcAg含量以精子细胞HBcAg指数(SHBcI)表示。乙型肝炎组和对照组SHBcI分别为(5.8777±8.9114)和(0.2800±0.1549),差异具有统计学意义;高病毒载量(HBV DNA≥106拷贝/ml)组SHBcI明显高于低病毒载量(HBV DNA<106拷贝/ml)组;24例拉米夫定治疗者,治疗后SHBcI明显下降,分别为(8.49±5.13)和(4.18±3.17)。结论该方法能够对精子细胞HBcAg定性兼定量进行诊断,该方法建立对于预测乙型肝炎父婴传播以及阻断治疗都将起到指导作用。

Objective To investigate and establish the special qualitative and quantitative analysis method of HBcAg in sperm ceils from hepatitis B patients by flow cytometry （FCM）. Methods Sperm was collected from sixty-two hepatiti B patients and the sperm cells were made into cellular suspension, then completely reacted with HBcAb monoclonal antibody immunologically marked by fluorescent,finally qualitatively and quantitatively measured by FCM. Results The sensitivity of this special qualitative analysis method of HBcAg in sperm cells from hepatitis/1 patients was 96.77% and the specificity was 100%, the positive pre- dictive value and the negative predictive value were both 100% , the accuracy were both 97.22%. The quantitation of HBcAg in sperm cells can be indicated by the sperm HBcAg index （SHBcI）. The SHBcI in hepatitis B and control group were （5. 8777 ± 8. 9114） and （0. 2800 ± 0. 1549）, respectively, with statistically significant difference The SHBcI in HBV DNA ≥ 10^6 copies/ml group was significantly higher than HBV DNA 〈 10^6 copies/ml group. The SHBeI had a distinct decline after lamivudine treatment （8.49 ± 5.13 ） vs （4.18 ± 3.17 ）. Conclusions This study set up a diagnosis method that could qualitatively and quantitatively measure HBeAg expression in sperm cells. It will play an important role in the prediction and interruption of father to infant transmission in patients with hepatitis B.