Zhang Aiying, Skog Sven, Wang Shengqi, Ke Yang, Zhang Yonghong, Li Kang, He Ellen, Li Ning
Beijing Institute of Hepatology, Beijing YouAn Hospital, Capital Medical University, Beijing 100069, China.
Sino-Swed Molecular Bio-Medicine Research Institute, Shenzhen 518057, China.
Sci Rep. 2016 Aug 16;6:31132. doi: 10.1038/srep31132.
The Lens culinaris agglutinin-reactive fraction of AFP (AFP-L3) is widely used to screen for hepatocellular carcinoma (HCC) in Japan and China. We developed a chemiluminescent protein microarray for determining the AFP-L3/AFP index (the ratio of AFP-L3 to total AFP, AFP-L3%) by fixing AFP-specific antibodies and Lens culinaris lectin on aldehyde-coated glass slides. Serum samples were tested for AFP using an enzyme-linked immunosorbent assay (ELISA) to validate the microarray. AFP-L3 was detected using Hotgen Biotech glycosyl capture spin column pretreatment technology and ELISA. When the AFP cut-off value was set to 20 ng/ml, the protein microarray displayed 89.74% sensitivity and 100% specificity for HCC diagnosis, and the ELISA displayed 87.17% sensitivity and 100% specificity. When the AFP-L3% cut-off value was set to 0.1, the protein microarray displayed 56.41% sensitivity and 100% specificity for HCC diagnosis, and the ELISA displayed 53.84% sensitivity and 100% specificity. The ROC curve for the HCC diagnosis showed that the AFP area under the ROC curve (AUC = 0.996; 95% CI: 0.986-1.005) was much higher than that of AFP-L3 (AUC = 0.857; 95% CI: 0.769-0.94) and AFP-L3% (AUC = 0.827; CI: 0.730-0.924). The microarray assay used in this study is a highly sensitive, accurate, and efficient assay for the determination of the AFP-L3%.
甲胎蛋白的豆凝集素反应性部分（AFP-L3）在日本和中国被广泛用于筛查肝细胞癌（HCC）。我们通过将甲胎蛋白特异性抗体和豆凝集素固定在醛包被的载玻片上，开发了一种用于测定AFP-L3/AFP指数（AFP-L3与总AFP的比值，即AFP-L3%）的化学发光蛋白质芯片。使用酶联免疫吸附测定法（ELISA）检测血清样本中的AFP，以验证该芯片。采用Hotgen Biotech糖基捕获旋转柱预处理技术和ELISA检测AFP-L3。当AFP临界值设定为20 ng/ml时，蛋白质芯片对HCC诊断的灵敏度为89.74%，特异性为100%，ELISA的灵敏度为87.17%，特异性为100%。当AFP-L3%临界值设定为0.1时，蛋白质芯片对HCC诊断的灵敏度为56.41%，特异性为100%，ELISA的灵敏度为53.84%，特异性为100%。HCC诊断的ROC曲线显示，AFP的ROC曲线下面积（AUC = 0.996；95% CI：0.986 - 1.005）远高于AFP-L3（AUC = 0.857；95% CI：0.769 - 0.94）和AFP-L3%（AUC = 0.827；CI：0.730 - 0.924）。本研究中使用的芯片检测法是一种用于测定AFP-L3%的高灵敏度、准确且高效的检测方法。
