双标记时间分辨荧光免疫分析检测AFP和AFP-IgM在HCC中的应用

目的建立可同时测定血清甲胎蛋白（AFP）和AFP免疫复合物（AFP-IgM）的快速、灵敏检测方法，探讨其在原发性肝细胞癌（HCC）诊断中的临床价值。方法利用时间分辨免疫荧光分析（TRFIA）技术，分别用Sm^3＋标记鼠抗人IgM单克隆抗体（M94181），Eu^3＋标记鼠抗人AFP单克隆抗体（M19301），建立双标记TRFIA（dual-TRFIA）。选择HCC患者118例和肝良性病变（BLD）患者123例（肝硬化60例，慢性肝炎63例），健康体格检查者286例。分别用该法检测血清AFP和AFP-IgM水平，并用受试者工作特征（ROC）曲线进行临床诊断特性分析。采用SASS 6．12软件进行统计学处理。结果dual-TRFIA法检测AFP和AFP-IgM的灵敏度分别为0．41μg／L，10．00U／L，与试剂盒[电化学发光免疫（ECLIA）AFP test和Hepa AFP-IC kit]相关性好（r=0．9987和0．9854，P均〈0．05）。AFP和AFP-IgM在HCC中的诊断灵敏度分别为46．61％（55／118），63．56％（75／118），诊断准确性分别为77．10％和77．40％。AFP和AFP-IgM联合检测可将灵敏度提高至72．88％（86／118，χ^2=9．45，P=0．02），诊断准确性提高至84．10％（χ^2=6．24，P=0．04）。结论AFP-IgM是HCC新型血清标志物，同时检测AFP和AFP-IgM能明显提高对HCC的诊断灵敏度和准确性，有利于肝癌的早期诊断。dual-TR-IMFA法同时检测AFP和AFP-IgM稳定可靠，可以满足临床使用需要。

Dual-label time-resolved immunofluorometric assay of serum AFP and AFP-IgM in patients with hepatocellular carcinoma

Objective Serum alpha-fetoprotein (AFP) is a widely used marker of hepatocellular carcinoma (HCC), although its diagnostic efficacy remains to be improved. Elevated levels of immunocomplex form of AFP-IgM in HCC patients have been reported. In this study a novel dual-labeled time-resolved immunofluorometric assay (dual-TRFIA) was developed to measure serum AFP and AFP-IgM levels simultaneously in order to establish a more effective serum maker detection method for HCC. Methods The dual-TRFIA for AFP-IgM or AFP was established by using Sm3+-labeled anti-human IgM monoclonal antibody (McAb) and Eu3+-labeled anti-AFP McAb as tracer antibodies. Serum AFP-IgM/AFP concentrations were measured in 118 patients with HCC, 60 patients with cirrhosis, 63 patients with chronic hepatitis and 286 healthy controls with the dual-TRFIA. The diagnostic efficacy was evaluated using receiver operating characteristic (ROC) curve. SAS 6.12 was used to analyze the data. Results The lowest detective concentrations for AFP and AFP-IgM were 0.41 ug/L and 10.00 U/L, respectively. AFP concentrations determined with duaI-TRFIA correlated well with those tested with electrochemiluminescenec immunoassay (ECLIA) (r= 0.9987, P<0.05). The sensitivity for diagnosing HCC with serum AFP and AFP-IgM was 46.61% (55/ 118) and 63.56% (75/118), respectively. The area under the ROC curve for AFP and AFP-IgM was 77.10% and 77.40%, respectively. However, the diagnostic sensitivity increased to 72.88% (86/118) (x2=9.45, P=0.02) and accuracy increased to 84.10% (x2=6.24, P=0.04) when the two markers were combined. Conclusions A highly sensitive and reliable dual-TRFIA for determination of serum AFP-IgM and AFP by a single test has been developed. It is suggested that combined measurement of serum AFP-IgM and AFP levels may improve the detection sensitivity of HCC.