合成多肽抗原检测原发性肝癌血清标志物ELISA方法的建立及应用  

Detection of Serum Markers of Primary Hepatocellular Carcinoma by ELISA Using Synthetic Polypeptide Antigen

作　　者：王文[1] 王锦红[1] 缪晓辉[2] Mark A Feitelson 戚中田[1] 

机构地区：[1]第二军医大学微生物学教研室上海市医学生物防护重点实验室,上海200433 [2]上海长征医院感染科,上海200003 [3]Department of Biology,College of Science and Technology,Temple University

出　　处：《中国生物制品学杂志》2009年第4期399-401,共3页Chinese Journal of Biologicals

基　　金：上海市重点学科建设项目(B901);第二军医大学青年启动基金项目(06QN03);艾滋病和病毒肝炎等重大传染病防治专项(200820001-017)

摘　　要：目的建立用合成多肽抗原检测原发性肝癌血清标志物的ELISA方法,并进行初步应用。方法利用筛选的5种与乙型肝炎病毒X抗原(HBx)诱导肝癌相关的细胞蛋白URG4、URG7、URG11、S15a和Sui1,作为原发性肝癌的筛查和早期诊断的血清标志物,根据上述蛋白的序列合成多肽(L4A/L4B、L7B、L11-1/L11-3/L11-4、L12A/L12B和Sui1A/Sui1B),作为检测抗原,建立检测相应抗体的间接ELISA法,并进行敏感性、特异性、精密性评价及临床应用。结果所建立的间接ELISA法敏感性为93.8%,特异性为97.9%,试验内和试验间变异系数分别为3.8%～5.6%和7.4%～10.3%;检测161份肝癌、肝硬化及乙型肝炎患者血清样本中,1种指标以上阳性检出率分别为90.4%、92.6%和61.7%,2种指标以上阳性检出率分别为78.6%、70.6%和48.3%;39份正常献血员血清中,1种指标以上阳性检出率为12.2%,2种指标以上阳性检出率为6.3%。结论所建立的ELISA法可作为目前的AFP和核磁共振诊断方法的有益补充,用于原发性肝癌高危人群的筛选和小肝癌的早期诊断。Objective To develop an ELISA method for detection of the serum markers of primary hepatocellular carcinoma （HCC）. Methods Five kinds of up-regulated cell proteins,i.e. URG4,URG7,URG11,S15a and Sui1,by HBx induction were served as serum markers of HCC in this paper. According to the sequences of above-mentioned proteins,polypeptides L4A / L4B,L7B,L11-1 / L11-3 / L11-4,L12A / L12B and Sui1A / Sui1B were synthesized and used as antigens for detection of the corresponding antibodies in sera by indirect ELISA. The developed ELISA method was evaluated for sensitivity,specificity and precision and used for detection of 161 samples from patients with HCC,liver cirthosis and hepatitis B and 39 from healthy blood donors. Results The sensitivity,specificity,inter-and intra-variation coefficients of developed ELISA method were 93. 8%,97. 9%,3. 8% ～ 5. 6% and 7. 4% ～ 10. 3% respectively. Of the serum samples from patients with HCC,liver cirthosis and hepatitis B,90. 4%,92. 6% and 61. 7% were positive for not less than 1 marker,and 78. 6%,70. 6% and 48. 3% for not less than 2 markers respectively. However,of the serum samples from healthy blood donors,12. 2% were positive for not less than 1 marker and 6. 3% for not less than 2 markers. Conclusion The developed ELISA may be used as an assistant diagnostic method of AFP and MRI for the screening of high risk population of HCC and the early diagnosis of small HCC.

关 键 词：肝癌 乙型肝炎病毒X抗原 多肽 血清标志物 ELISA 

分 类 号：R446[医药卫生—诊断学]