Overexpression and purification of a recombinant chimeric HIV type 2 HIV type 1 envelope peptide and application in an accelerated immunobased HIV type 1/2 antibody detection system (AIBS): A new rapid serological screening assay
Overexpression and purification of a recombinant chimeric HIV type 2 HIV type 1 envelope peptide and application in an accelerated immunobased HIV type 1/2 antibody detection system (AIBS): A new rapid serological screening assay
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DOI：
10.1089/aid.1996.12.1081
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发表时间：
1996-08-10
影响因子：
1.5
通讯作者：
Pitner, F
中科院分区：
文献类型：
--
作者：
Ecker, B;Vollenhofer, S;Pitner, F
构建了包含HIV-2 gp36免疫优势区和HIV-1 gp41免疫优势区的HIV-2/HIV-1包膜嵌合序列，并在大肠杆菌中高效表达。将重组产物(rp21/18)纯化后应用于一种新的抗体筛选方法，新原理的设计特点是：(1)总检测时间约为30min：(2)检测过程包括三个操作步骤；(3)本试验具有可靠的灵敏度和特异度，将稀释的血清样品和G蛋白-辣根过氧化物酶结合物同时加入包被的(杂合抗原HIV-1/2)和封闭的微孔板中，使血清样品与G蛋白-辣根过氧化物酶的批量孵育省去了经典的ELISA5步中通常需要的两个操作步骤。用市售血清转换板和来自血库的随机阴性血清样本对AIBS进行评估，血清转换结果表明AIBS对ELISA和第三代检测具有同等的敏感性，对5012名献血者(奥地利维也纳红十字会)进行了特异性测定，献血者群体的重复反应率为0.02%。AIBS代表了一个通用的免疫测定系统(不仅仅是HIV抗体)，对AIBS进行HIV-1/2筛查评估的整个AIBS检测过程，包括结果报告，可以由几个商业系统自动执行。根据这些系统，AIBS在实验室或血库中可能有用，因为这些实验室或血库同时进行了高容量和低容量检测。
A chimeric HIV-2/HIV-1 envelope sequence containing an immunodominant region of HIV-2 gp36 and the corresponding region of HIV-1 gp41 was constructed and overexpressed in Escherichia coli. The recombinant product (rp21/18) was purified and applied in a novel antibody-screening assay, Characteristics in the design of this new principle are as follows: (1) the overall assay time is about 30 min; (2) the assay procedure includes three manipulation steps; and (3) the test shows a reliable performance with respect to sensitivity and specificity, The diluted serum sample and the protein G-horseradish peroxidase conjugate are added simultaneously into a coated (hybrid antigen HIV-1/2) and blocked microtiter plate well, The in-batch incubation of serum sample with protein G-horseradish peroxidase saves two manipulation steps that are normally necessary in the five-step procedure of a classical ELISA. AIBS was evaluated with commercially available seroconversion panels and with random negative serum samples from a blood bank, Seroconversion results demonstrated that AIBS has equivalent sensitivity to ELISAs and the third generation assays, The specificity was determined on a total blood donor population of 5012 (Red Cross Vienna, Austria), The repeat reactive rates for donor population were 0.02%. AIBS represents a general immunometric system (not only HIV antibodies), The entire assay procedure of AIBS evaluated for HIV-1/2 screening, including result reporting, can be performed automatically by several commercially available systems. Depending on these systems AIBS is potentially useful in laboratories or blood banks that have both high- and low-volume testing.