检测人博卡病毒抗体间接酶联免疫吸附法的建立及临床应用  

Establishment and clinical use of indirect ELISA for detection of human bocavirus （HBoV） antibody

作　　者：林峰[1] 滕灵方[2] 郑敏巧[3] 郑昌华[1] 吴锋[3] 黄恩佩[2] 侯建毅[2] 

机构地区：[1]温州医学院附属温岭医院儿科,317500 [2]温州医学院附属温岭医院生物芯片研究中心,317500 [3]温州医学院附属温岭医院检验科,317500

出　　处：《中华生物医学工程杂志》2010年第4期359-362,共4页Chinese Journal of Biomedical Engineering

基　　金：基金项目：国家自然科学基金（30771913）;国家高技术研究发展“863”计划基金（2007AA022463）

摘　　要：目的建立间接酶联免疫吸附法（ELISA）检测人博卡病毒（HBoV）抗体，探讨其临床应用的可行性。方法以重组表达的HBoV融合蛋白VP2作为包被抗原，建立检测HBoV抗体的间接ELISA，优化该检测方法的最佳条件，观察其精密度、敏感度和特异性，并与荧光定量PCR方法对比检测40份临床样本，同时采用免疫印迹法（Westernblot）确认其符合率。结果所建立的间接ELISA最佳抗原包被浓度为2mg／L，酶标二抗工作浓度为1：5000，血清标本最佳稀释度为1：200。该方法平均批内变异系数为6．87％，平均批间变异系数为4．67％。40份临床样本间接ELISA检测结果与荧光定量PCR及免疫印迹结果一致，符合率100％。结论利用VP2融合蛋白作为抗原，建立的检测血清HBoV抗体间接ELISA方法特异性强、重复性好，可用于HBoV抗体的定量和定性检测。Objective To establish indirect ELISA for detection of HBoV antibody and to evaluate the feasibility of its clinical use. Methods Using the HBoV fusion protein VP2 generated by recombinant expression as envelope antigen, indirect ELISA method for HBoV antibody detection was established. The optimal condition of the test was set, and precision, sensitivity and specificity were examined. The findings of the test were compared with those using quantitative fluorescence PCR in 40 clinical samples, and were validated by Western blot to evaluate the consistency between each other. Results The optimal concentration of envelop antigen, enzyme-labeled secondary antigen and the serum dilution was 2 mg/L, 1 ： 5000 and 1：200, respectively. The coefficients of intra-assay and inter-assay variations were 6.87% and 4.67%. The findings of this test in 40 clinical samples were 100% consistent with those by quantitative fluorescence PCR and Western blot. Conclusion The new indirect ELISA method for HBoV antibody detection using HBoV fusion protein VP2 as envelope antigen is highly specific, reproducible and useful for quantitative or qualitative detection of HBoV antibody.

关 键 词：博卡病毒属 抗体 酶联免疫吸附测定 病毒融合蛋白质类 临床实验室技术 

分 类 号：R446[医药卫生—诊断学]