College of Pharmacy, Chungbuk National University, Chungbuk 361-763, Korea.
Microbiol Immunol. 2012 Jun;56(6):372-7. doi: 10.1111/j.1348-0421.2012.00455.x.
We prepared mAb specific to the H1N1 2009 virus (H1N1 2009) to facilitate development of an RDT with enhanced sensitivity and specificity. Among these antibodies, we identified two clones--hybridomas 1H7E1 and 3A3H7-that specifically bound to H1N1 2009 (non-seasonal) and were very suitable for application to a diagnostic kit. The affinity constants (K(a)) of 1H7E1 and 3A3H7 were 1.10 × 10(10) and 2.35 × 10(10), respectively. To identify the antibodies, we performed ELISA and immunoblot analyses and found that 1H7E1 recognized a conformational epitope of HA while 3A3H7 recognized a linear epitope. In clinical evaluations using specimens from 215 patients, a lateral flow rapid testing kit comprising these mAb showed a sensitivity of 81.5% (75/92) and a specificity of 96.7% (119/123). Results using the RDT kit were well correlated with conventional RT-PCR methods as commonly and commercially used. Based on our findings, we believe that use of these mAb with a rapid evaluation kit could serve as a good diagnostic tool for H1N1 2009.
我们制备了针对 H1N1 2009 病毒（H1N1 2009）的 mAb，以促进开发具有更高灵敏度和特异性的 RDT。在这些抗体中，我们鉴定出两个克隆——杂交瘤 1H7E1 和 3A3H7——它们特异性结合 H1N1 2009（非季节性），非常适合应用于诊断试剂盒。1H7E1 和 3A3H7 的亲和力常数（K(a)）分别为 1.10×10(10)和 2.35×10(10)。为了鉴定这些抗体，我们进行了 ELISA 和免疫印迹分析，发现 1H7E1 识别 HA 的构象表位，而 3A3H7 识别线性表位。在使用来自 215 名患者的标本进行的临床评估中，包含这些 mAb 的侧向流动快速检测试剂盒的灵敏度为 81.5%（75/92），特异性为 96.7%（119/123）。使用 RDT 试剂盒的结果与常规 RT-PCR 方法高度相关，这些方法通常在临床上使用。基于我们的研究结果，我们认为使用这些 mAb 与快速评估试剂盒一起可以作为一种良好的诊断 H1N1 2009 的工具。
