Development and implementation of an international proficiency testing program for a neutralizing antibody assay for HIV-1 in TZM-bl cells.
Development and implementation of an international proficiency testing program for a neutralizing antibody assay for HIV-1 in TZM-bl cells.
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DOI：
10.1016/j.jim.2011.09.007
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发表时间：
2012-01-31
影响因子：
2.2
通讯作者：
Sarzotti-Kelsoe, Marcella
中科院分区：
文献类型：
--
作者：
Todd, Christopher A.;Greene, Kelli M.;Yu, Xuesong;Ozaki, Daniel A.;Gao, Hongmei;Huang, Yunda;Wang, Maggie;Li, Gary;Brown, Ronald;Wood, Blake;D'Souza, M. Patricia;Gilbert, Peter;Montefiori, David C.;Sarzotti-Kelsoe, Marcella
最近检测技术的进步使艾滋病毒-1中和抗体的检测方法有了重大改进。在TZM-bl(JC53bl-13)细胞中进行的荧光素酶报告基因分析已经得到优化和验证。由于这种检测方法已被世界各地的多家实验室采用，因此开发了一项外部熟练程度测试计划，以确保为HIV/AIDS疫苗临床试验执行这种中和抗体检测的实验室之间的数据等价性。该计划通过进行三轮独立测试进行了优化，从第一轮到第三轮的严格程度有所提高。参与的国内和国际实验室的结果每一轮都有所改善，因为有助于分析间可变性的因素被确定并降至最低。增加协议的关键因素是实验室之间的经验和试剂的标准化。使用基于三轮优化测试的模拟程序开发了统计合格规则，其中，如果30个ID50值中至少有25个在接受范围内，实验室就合格。这确保了参与的实验室不会有超过20%的风险，即当它应该符合模拟程序的定义时，它没有获得资格。确定了五个有经验的参考实验室，并对一系列标准化试剂进行了测试，以得出合格-不合格标准的接受范围。该标准化水平测试计划是第一个可用于评估和记录执行HIV-1中和抗体检测的实验室的测试等价性的标准水平测试计划，并可能为未来开发其他测试平台的水平测试计划提供指导。
Recent advances in assay technology have led to major improvements in how HIV-1 neutralizing antibodies are measured. A luciferase reporter gene assay performed in TZM-bl (JC53bl-13) cells has been optimized and validated. Because this assay has been adopted by multiple laboratories worldwide, an external proficiency testing program was developed to ensure data equivalency across laboratories performing this neutralizing antibody assay for HIV/AIDS vaccine clinical trials. The program was optimized by conducting three independent rounds of testing, with an increased level of stringency from the first to third round. Results from the participating domestic and international laboratories improved each round as factors that contributed to inter-assay variability were identified and minimized. Key contributors to increased agreement were experience among laboratories and standardization of reagents. A statistical qualification rule was developed using a simulation procedure based on the three optimization rounds of testing, where a laboratory qualifies if at least 25 of the 30 ID50 values lie within the acceptance ranges. This ensures no more than a 20% risk that a participating laboratory fails to qualify when it should, as defined by the simulation procedure. Five experienced reference laboratories were identified and tested a series of standardized reagents to derive the acceptance ranges for pass–fail criteria. This Standardized Proficiency Testing Program is the first available for the evaluation and documentation of assay equivalency for laboratories performing HIV-1 neutralizing antibody assays and may provide guidance for the development of future proficiency testing programs for other assay platforms.
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DOI：
10.1016/j.bbrc.2006.07.163
发表时间：
2006-09-29
影响因子：
3.1
作者：
Choudhry, Vidita;Zhang, Mei-Yun;Dimitrov, Dimiter S.
通讯作者：
Dimitrov, Dimiter S.
国际艾滋病毒中和试验比较网络：NeutNet 报告。
影响因子：
3.7
作者：
Fenyo, Eva Maria;Heath, Alan;Dispinseri, Stefania;Holmes, Harvey;Lusso, Paolo;Zolla-Pazner, Susan;Donners, Helen;Heyndrickx, Leo;Alcami, Jose;Bongertz, Vera;Jassoy, Christian;Malnati, Mauro;Montefiori, David;Moog, Christiane;Morris, Lynn;Osmanov, Saladin;Polonis, Victoria;Sattentau, Quentin;Schuitemaker, Hanneke;Sutthent, Ruengpung;Wrin, Terri;Scarlatti, Gabriella
通讯作者：
Scarlatti, Gabriella
影响因子：
6.4
作者：
DSouza, MP;Livnat, D;Katinger, H
通讯作者：
Katinger, H
影响因子：
15.8
作者：
Sarzotti-Kelsoe M;Cox J;Cleland N;Denny T;Hural J;Needham L;Ozaki D;Rodriguez-Chavez IR;Stevens G;Stiles T;Tarragona-Fiol T;Simkins A
通讯作者：
Simkins A
影响因子：
5.4
作者：
Li, Ming;Salazar-Gonzalez, Jesus F.;Montefiori, David C.
通讯作者：
Montefiori, David C.