基于SPRi生物芯片平台的HIV-1感染快速检测方法的建立及其应用  

The establishment and use of a technique for rapid detection of HIV-1 infection using an SPRi biosensor system

作　　者：汪海波[1] 杨泽[1] 陈新彬 廉国胜[1] 史咏梅[1] 冯子力[1] 伍碧梅[1] 莫秋华[1] WANG Hai-bo;YANG Ze;CHEN Xin-bin;LIAN Guo-sheng;SHI Yong-mei;FENG Zi-li;WU Bi-mei;MO Oiu-hua(State Key Laboratory for Detection of Emerging Infectious Diseases,Zhuhai International Travel Healthcare Center,Zhuhai 519020,Guangdong,China)

机构地区：[1]新发传染病检测重点实验室珠海国际旅行卫生保健中心,广东珠海519020

出　　处：《中国病原生物学杂志》2018年第8期823-826,共4页Journal of Pathogen Biology

基　　金：广东省科技计划项目(No.2015A020211004);国家质检总局科技计划项目(No.2016IK302)

摘　　要：目的利用表面等离子共振成像(SPRi)生物芯片平台建立HIV-1感染快速检测方法并将其应用于临床标本的检测。方法通过文献分析比较筛选出最适检测蛋白,再通过梯度稀释法确定其适宜检测浓度,建立HIV-1感染检测方法并进行优化,然后用已知标本(HIV-1阳性血清,HBV阳性血清,HCV阳性血清及健康人血清)进行检测以确定该方法的特异性;与商业化ELISA检测试剂同步用于临床标本的检测,比较检测结果的一致性。结果当检测蛋白浓度低于0.16mg/ml时,无阳性信号出现;当检测蛋白浓度≥0.16mg/ml时均有阳性信号出现且呈现线性增强趋势。从降低检测成本和减少高浓度时可能出现非特异性结合的情况考虑,最终将检测蛋白的最适浓度确定为0.16mg/ml。用优化的HIV-1检测方法检测已知标本,只有HIV-1阳性血清出现阳性信号,HBV阳性血清、HCV阳性血清及健康人血清时均无阳性信号。临床标本检测表明,该方法和商业化ELISA试剂盒,均能检出同一份HIV-1阳性标本,一致性为100%。结论基于SPRi平台建立的HIV-1快速检测法具有特异、准确、快速、简便等优点,可用于HIV-1感染检测。Objectives To establish a method for rapid detection of HIV-1 infection using an SPRi biosensor system and to evaluate its performance in a clinical setting. Methods A detection antigen was screened through an analysis of the literature and its concentration was determined using serial dilution.Then several rounds of optimization were performed to establish a method for rapid detection of HIV-1 infection.The specificity of this method was evaluated using confirmed samples（including sera from HIV-1-positive,HBV-positive,and HCV-positive patients and healthy donors）.The concordance between this method and a commercial ELISA assay was also evaluated. Results No positive signal appeared when the concentration of the detection antigen was below 0.16 mg/ml.A positive signal appeared and intensified as the concentration of the detection antigen increased.The optimized concentration of the detection antigen was set at0.16 mg/mL based on comprehensive considerations,including controlling costs and decreasing non-specific binding at high antigen concentrations.Results from confirmed samples indicated that the method was highly specific to HIV-1 and did not cross-react with HBV or HCV.Clinical use indicated that the method was as accurate as a commercial ELISA assay,with a concordance of 100%. Conclusion A method of HIV-1 detection based on an SPRi biosensor system was specific,accurate,rapid,and simple,and this method could be used to detect HIV-1 infection.

关 键 词：SPRi 生物芯片 HIV-1 快速检测 

分 类 号：R373.9[医药卫生—病原生物学]