丙型肝炎病毒非结构蛋白NS2核酸适配子的功能鉴定  

Functional identification of hepatitis C virus non-structural protein NS2 aptamers

作　　者：许少英[1] 刘凯媚[1] 张婷[1] 李仲平[1] XU Shao-ying;LIU Kai-mei;ZHANG Ting;LI Zhong-ping(Guangzhou Blood Center,Guangzhou Key Medical Laboratory(Blood Safety Key Laboratory),Guangzhou,Guangdong 510095,China)

机构地区：[1]广州血液中心,广州市医学重点实验室(血液安全重点实验室),广东广州510095

出　　处：《热带医学杂志》2020年第7期903-906,共4页Journal of Tropical Medicine

基　　金：广州市医学重点学科建设项目(血液安全重点实验室)。

摘　　要：目的对丙型肝炎病毒非结构蛋白NS2核酸适配子功能进行鉴定。方法应用ELISA方法检测适配子与HCV-NS2蛋白的亲和力,检测适配子与HCV阳性血清的结合能力和适配子与不同比例稀释的HCV阳性血清的结合能力,并与HCV阴性血清比较。结果筛选得到4条适配子序列A1、A2、A3、A4。4条适配子与HCV-NS2蛋白的亲和力检测吸光度值分别为:A1:0.263、A2:0.515、A3:0.427、A4:0.339,均高于阴性对照吸光度值0.001,差异有统计学意义(P≤0.001);4条适配子均与HCV-NS2蛋白有较高的亲和力,其中,适配子A2与HCV-NS2蛋白的亲和力最高。适配子A2与10人份不同人来源的HCV阳性献血者血清(P1-P10)的结合吸光度分别为0.157、0.145、0.165、0.140、0.241、0.129、0.137、0.222、0.138、0.136,高于其与阴性血清结合吸光度0.006,差异有统计学意义(P≤0.001)。HCV-NS2与1∶0、1∶1、1∶2稀释的HCV阳性血清结合吸光度分别为0.223、0.151、0.093,均高于其与阴性血清结合吸光度0.005,差异有统计学意义(P≤0.001);HCV-NS2适配子与不同比例稀释的HCV阳性血清的结合呈剂量依赖性。结论筛选得到的HCV-NS2适配子功能鉴定表明其具有较强的应用价值。Objective To identify the aptamer of the non-structural protein NS2 of hepatitis C virus.Methods ELISA methods was used to detect the affinity between the aptamer and HCV-NS2 protein.The binding capacity of aptamers to HCV positive sera and the binding capacity of aptamers to HCV positive sera diluted in different proportions were detected and compared with HCV negative sera.Results Four aptamer sequences A1,A2,A3 and A4 were obtained.The absorbance values of four aptamers with HCV-NS2 protein were A1:0.263,A2:0.515,A3:0.427,and A4:0.339,which were significantly higher than those of negative control(P≤0.001).All of the four aptamers had high affinity with HCV-NS2 protein;among which,A2 had the highest affinity with HCV-NS2 protein.The binding absorbances of aptamer A2 with sera of 10 HCV positive donors(P1-P10)from different sources were 0.157,0.145,0.165,0.140,0.241,0.129,0.137,0.222,0.138 and 0.136,respectively,which were higher than those of negative sera(P<0.001).The binding absorbances of aptamer A2 with 1∶0,1∶1 and 1∶2 diluted HCV positive sera were 0.223,0.151 and 0.093,respectively,which were significantly higher than those with negative serum(P≤0.001),and showing a dose-dependent manner.Conclusion According to the results of functional identification,aptamer of HCV-NS2 had application value.

关 键 词：丙型肝炎病毒 NS2蛋白 适配子 功能 

分 类 号：R512.63[医药卫生—内科学]