目的:采用时间分辨荧光免疫技术(TrFIA)建立高灵敏的人附睾分泌蛋白4(HE4)的检测方法.方法:双抗体夹心法建立HE4-TrFIA,并对该试剂的各项性能指标进行评价.结果:该法的检测范围为1.08pmol/L ~2000pmol/L,灵敏度为1.08pmol/L,HE4的回收率95.6%,批内和批间变异系数分别3.6%~7.3%,4.6%~10.2%.与CA125交叉反应率0.01%.破坏试验表明,试剂在37℃可稳定7d.62例卵巢癌患者和60例健康人血清,用该试剂盒测得卵巢癌患者血清HE4浓度显著高于健康人(P<0.001).该试剂盒检测结果与进口HE4 kit检测结果r为0.951.结论:自建的HE4-TrFIA是一种敏感度高、特异性强、准确度高的适用方法,具有良好的临床应用前景.%Objective To develop a time-resolved fluoroimmunoassay (TrFIA) with high sensitivity for detecting human epididy-mis protein 4(HE4) in serum. Methods Sandwich TrFIA kit for HE4 detection was developed using monoclonal antibody of anti-HE4 and its methodology was evaluated. Results HE4-TrFIA kit was capable of detecting HE4 within the range of 1.08-2000 pmol/L with the sensitivity of 1.08 pmol/L and without cross-reactivity with CA125. The intra-assay and inter-assay coefficients of variation were 3.6% -7.3% and 4.6% -10.2% , respectively. The prepared HE4-TrFIA reagent could withstand preservation at 37℃ for 7 days. Serum samples were collected from 62 ovary carcinoma and 60 normal healthy controls. HE4 were significantly higher in ovary carcinoma patients compared with controls (P<0.001). The correlation coefficient of the detection results between this kit and commercially a-vailable FUJIREBIO kit for 122 blood samples was 0.951. Conclusion The prepared TrFIA, possess high sensitivity ,specificity and precision is a suitable method for clinical spplication.
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