Johnson A J, Martin D A, Karabatsos N, Roehrig J T
Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Public Health Service, U.S. Department of Health and Human Services, Fort Collins, Colorado 80522, USA.
J Clin Microbiol. 2000 May;38(5):1827-31. doi: 10.1128/JCM.38.5.1827-1831.2000.
Monoclonal antibody (MAb)-based capture enzyme-linked immunosorbent assays (ELISAs) for the detection of anti-arboviral immunoglobulin G (IgG ELISAs) were developed for a comprehensive array of medically important arboviruses from the Alphavirus, Flavivirus, and Bunyavirus genera. Tests were optimized and standardized so that maximum homology could be maintained among working parameters for the different viral agents, enabling a wide range of viruses to be easily tested for at one time. MAbs were screened for suitability as capture vehicles for antigens from the three genera. The final test configuration utilized group-reactive MAbs eastern equine encephalitis virus 1A4B-6, dengue 2 virus 4G2, and La Crosse encephalitis virus 10G5.4 to capture the specific inactivated viral antigens. Serum IgG was detected by using alkaline phosphatase-conjugated anti-human IgG (Fc portion). A dilution of 1:400 was chosen as the universal screening serum dilution, with endpoint titrations of serum samples testing positive eliminating occasional false-positive results. IgG ELISA results correlated with those of the standard plaque-reduction neutralization assays. As expected, some test cross-reactivity was encountered within the individual genera, and tests were interpreted within the context of these reactions. The tests were standardized for laboratory diagnosis of arboviral infections, with the intent that they be used in tandem with the corresponding IgM antibody-capture ELISAs.
基于单克隆抗体（MAb）的捕获酶联免疫吸附测定（ELISA）用于检测抗虫媒病毒免疫球蛋白G（IgG ELISA），已针对α病毒属、黄病毒属和布尼亚病毒属中一系列具有重要医学意义的虫媒病毒开发出来。对检测进行了优化和标准化，以便在不同病毒制剂的工作参数之间保持最大同源性，从而能够一次轻松检测多种病毒。筛选单克隆抗体作为来自这三个属的抗原的捕获载体。最终的检测配置使用了组反应性单克隆抗体东部马脑炎病毒1A4B - 6、登革2病毒4G2和拉克罗斯脑炎病毒10G5.4来捕获特定的灭活病毒抗原。通过使用碱性磷酸酶偶联的抗人IgG（Fc段）检测血清IgG。选择1:400的稀释度作为通用的筛查血清稀释度，对检测呈阳性的血清样本进行终点滴定以消除偶尔出现的假阳性结果。IgG ELISA结果与标准蚀斑减少中和试验的结果相关。正如预期的那样，在各个属内遇到了一些检测交叉反应，并在这些反应的背景下解释检测结果。这些检测针对虫媒病毒感染的实验室诊断进行了标准化，旨在与相应的IgM抗体捕获ELISA联合使用。
