Qiu Li-Wen, Di Biao, Wen Kun, Wang Xin-shuai, Liang Wei-hua, Wang Ya-di, Pan Yu-xian, Wang Ming, Ding Yan-qing, Che Xiao-yan
Clinical Laboratory, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
Clin Vaccine Immunol. 2009 Jan;16(1):88-95. doi: 10.1128/CVI.00212-08. Epub 2008 Nov 19.
The dengue virus (DENV) has four distinct serotypes (DENV1, DENV2, DENV3, and DENV4) that require differentiation for effective prevention of morbid diseases. The recently developed DENV1-specific NS1 antigen capture enzyme-linked immunosorbent assay (ELISA) based on the monoclonal antibodies (MAbs) that recognize distinct epitopes on nonstructural protein 1 (NS1) of a specific DENV serotype is convenient and cost-effective, but assays have not yet been developed for DENV serotypes 2 to 4. This paper describes the development and validation of a DENV2-specific NS1 antigen capture ELISA by selection and optimization of the pair of well-characterized MAbs that recognized epitopes specific for DENV2 NS1 from a large panel of MAbs. The DENV2 NS1 ELISA displayed exclusive sensitivity with the DENV2 serotype and did not cross-react with the other three DENV serotypes. The sensitivity and specificity of the DENV2 NS1 ELISA were 83.3% (25/30) and 100% (504/504) when used to test 30 acute-phase serum samples from patients infected with DENV2 identified by virus isolation or reverse transcription-PCR serotyping and 504 serum samples from healthy individuals, respectively. The specificity of this assay was also evaluated using a panel of serum samples which were positive for DENV1, other flaviviruses, and nonflaviviruses; no cross-reactions were observed in these clinical samples. The DENV2 NS1 ELISA was eightfold more sensitive than a commercially available serotype-cross-reactive NS1 ELISA (Panbio Diagnostics, Brisbane, Australia) when the two assays were used to test the DENV2-infected cell culture supernatants in parallel. The Panbio NS1 ELISA displayed variation in sensitivity between DENV serotypes. The DENV2-specific NS1 antigen capture ELISA can be used as a tool for the rapid identification of DENV2 infections.
登革病毒（DENV）有四种不同的血清型（DENV1、DENV2、DENV3和DENV4），为有效预防发病，需要对它们进行区分。最近基于单克隆抗体（MAb）开发的DENV1特异性NS1抗原捕获酶联免疫吸附测定（ELISA），该单克隆抗体可识别特定DENV血清型非结构蛋白1（NS1）上的不同表位，这种方法方便且具有成本效益，但尚未开发出针对DENV 2至4血清型的检测方法。本文描述了通过从大量单克隆抗体中选择和优化一对特性明确的单克隆抗体来开发和验证DENV2特异性NS1抗原捕获ELISA，这对单克隆抗体可识别DENV2 NS1特有的表位。DENV2 NS1 ELISA对DENV2血清型具有独特的敏感性，且不与其他三种DENV血清型发生交叉反应。当用于检测30份经病毒分离或逆转录 - PCR血清分型鉴定为感染DENV2的患者急性期血清样本和504份健康个体血清样本时，DENV2 NS1 ELISA的敏感性和特异性分别为83.3%（25/30）和100%（504/504）。还使用一组对DENV1、其他黄病毒和非黄病毒呈阳性的血清样本评估了该检测方法的特异性；在这些临床样本中未观察到交叉反应。当两种检测方法并行检测感染DENV2的细胞培养上清液时，DENV2 NS1 ELISA的敏感性比市售的血清型交叉反应NS1 ELISA（澳大利亚布里斯班的Panbio Diagnostics公司生产）高八倍。Panbio NS1 ELISA在不同DENV血清型之间的敏感性存在差异。DENV2特异性NS1抗原捕获ELISA可作为快速鉴定DENV2感染的工具。
