Department of Immunology, Duke University Medical Center, Durham, NC, United States; Department of Surgery, Duke University Medical Center, Durham, NC, United States.
Vaccine Research Center, National Institutes of Health (NIH), Bethesda, MD, United States.
J Immunol Methods. 2014 Jul;409:131-46. doi: 10.1016/j.jim.2013.11.022. Epub 2013 Dec 1.
The TZM-bl assay measures antibody-mediated neutralization of HIV-1 as a function of reductions in HIV-1 Tat-regulated firefly luciferase (Luc) reporter gene expression after a single round of infection with Env-pseudotyped viruses. This assay has become the main endpoint neutralization assay used for the assessment of pre-clinical and clinical trial samples by a growing number of laboratories worldwide. Here we present the results of the formal optimization and validation of the TZM-bl assay, performed in compliance with Good Clinical Laboratory Practice (GCLP) guidelines. The assay was evaluated for specificity, accuracy, precision, limits of detection and quantitation, linearity, range and robustness. The validated manual TZM-bl assay was also adapted, optimized and qualified to an automated 384-well format.
TZM-bl 测定法可测量抗体介导的 HIV-1 中和作用，其作为在单次 Env 假型病毒感染后 HIV-1 Tat 调控的萤火虫荧光素酶（Luc）报告基因表达减少的函数。该测定法已成为全球越来越多的实验室用于评估临床前和临床试验样品的主要终点中和测定法。在这里，我们根据良好临床实验室实践（GCLP）指南，介绍了 TZM-bl 测定法的正式优化和验证结果。该测定法的特异性、准确性、精密度、检测限和定量限、线性、范围和稳健性进行了评估。经过验证的手动 TZM-bl 测定法也被改编、优化并适用于自动化 384 孔格式。
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