脱-γ-羧基凝血酶原的Ba-蛋白芯片法的建立和应用  

Establishment and Application of Ba-Protein Microarray of Des-γ-carboxy-prothrombin

作　　者：李康[1,2] 张爱英[2,3] 张贺[4] 赵元顺[1,2] 张永宏[1,2] 李宁[1,2,3] 

机构地区：[1]首都医科大学附属北京佑安医院,北京100069 [2]传染病相关生物标志物北京市重点实验室,北京100069 [3]首都医科大学附属北京佑安医院北京市肝病研究所,北京100069 [4]军事医学科学院放射与辐射医学研究所生物技术研究室,北京100850

出　　处：《现代生物医学进展》2015年第33期6415-6419,共5页Progress in Modern Biomedicine

基　　金：国家科技部国际科技合作专项(S2012ZR0153);"十二五"传染病重大科技专项(2013X10004001-001-003);北京市科学技术委员会重点项目(D131100005313004;D131100005313005);首都医科大学重点实验室基金项目(2013GYGA03);重大传染病防治协同创新中心资助

摘　　要：目的:研发建立一种快速、特异性高、低成本的检测血清中脱-γ-羧基凝血酶原(DCP)的蛋白芯片,并通过检测DCP阳性肝细胞肝癌(HCC)血清临床样本对其进行评价。方法:首先采用柠檬酸钡吸附沉淀法去除血清中正常凝血酶原,然后通过双抗体夹心法蛋白芯片检测血清中DCP含量。通过Ba-蛋白芯片法检测41例DCP阳性HCC患者血清和66例健康人血清,进行盲法验证。结果:通过Ecarin-显色多肽底物法证实柠檬酸钡吸附沉淀法能够完全除血清中正常凝血酶原,根据"平均数+3×标准差"公式设置蛋白芯片的Cut-off值作为阳性判定标准,Ba-蛋白芯片法检测DCP阳性HCC血清的灵敏度为95.1%(39/41),特异度为100%(66/66)。结论:成功建立检测血清中DCP浓度的Ba-蛋白芯片法,为多种标志物联合检测HCC的蛋白芯片的研发提供候选血清标志物。Objective： To develop a rapid, highly specific, and low-cost protein microarray for detecting des-γ-carboxy- prothrombin （DCP）, and to evaluate it through diagnosis of DCP-positive hepatocellular carcinoma （HCC） clinical samples. Methods： Firstly, the removal of normal prothrombin purified by adsorption and precipitation method with barium citrate in plasma, and second using double-antibody sandwich method, the protein microarray were prepared for detecting DCP level. 41 DCP positive serum samples from patients with HCC and 66 serum samples from healthy human were detected with Ba-protein microarray in the blind experiment. Results： Ecarin-chromogenic peptide substrate assay indicated that normal prothrombin was completely eliminated by barium citrate adsorption and precipitation method. According to ＂average ＋ 3 × standard deviation＂ formula setting Cut-offvalue of protein chip as a positive criteria, the sensitivity ofBa-protein microarray was 95.1% （39/41）, and its specificity was 100 % （66/66）. Conclusions： The Ba-protein microarray for detecting DCP in serum was successfully established, and provided one candidate biomarker to research multiple biomarkers protein microarray for association detection of HCC.

关 键 词：Ba-蛋白芯片法 脱-Γ-羧基凝血酶原 肝细胞肝癌 血清标志物 

分 类 号：R730.3[医药卫生—肿瘤] R735.7[医药卫生—临床医学]