Tang Yin-Liang, Chiu Chien-Yu, Lin Chun-Yu, Huang Chung-Hao, Chen Yen-Hsu, Destura Raul V, Chao Day-Yu, Wu Han-Chung
Graduate Institute of Life Sciences, National Defense Medical Center, Taipei 114, Taiwan.
Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan.
Int J Mol Sci. 2015 Nov 24;16(11):27850-64. doi: 10.3390/ijms161126069.
Dengue virus (DENV) infection is currently at pandemic levels, with populations in tropical and subtropical regions at greatest risk of infection. Early diagnosis and management remain the cornerstone for good clinical outcomes, thus efficient and accurate diagnostic technology in the early stage of the disease is urgently needed. Serotype-specific monoclonal antibodies (mAbs) against the DENV1 nonstructural protein 1 (NS1), DA12-4, DA13-2, and DA15-3, which were recently generated using the hybridoma technique, are suitable for use in diagnostic platforms. Immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (ELISA) and Western blot analysis further confirmed the serotype specificity of these three monoclonal antibodies. The ELISA-based diagnostic platform was established using the combination of two highly sensitive mAbs (DA15-3 and DB20-6). The same combination was also used for the flow cytometry-based diagnostic platform. We report here the detection limits of flow cytometry-based and ELISA-based diagnostic platforms using these mAbs to be 0.1 and 1 ng/mL, respectively. The collected clinical patient serum samples were also assayed by these two serotyping diagnostic platforms. The sensitivity and specificity for detecting NS1 protein of DENV1 are 90% and 96%, respectively. The accuracy of our platform for testing clinical samples is more advanced than that of the two commercial NS1 diagnostic platforms. In conclusion, our platforms are suitable for the early detection of NS1 protein in DENV1 infected patients.
登革病毒（DENV）感染目前处于大流行水平，热带和亚热带地区的人群感染风险最高。早期诊断和管理仍然是取得良好临床结果的基石，因此迫切需要疾病早期高效、准确的诊断技术。最近利用杂交瘤技术产生的针对登革病毒1型非结构蛋白1（NS1）的血清型特异性单克隆抗体（mAb）DA12 - 4、DA13 - 2和DA15 - 3适用于诊断平台。免疫荧光测定（IFA）、酶联免疫吸附测定（ELISA）和蛋白质印迹分析进一步证实了这三种单克隆抗体的血清型特异性。基于ELISA的诊断平台是使用两种高灵敏度单克隆抗体（DA15 - 3和DB20 - 6）组合建立的。相同的组合也用于基于流式细胞术的诊断平台。我们在此报告，使用这些单克隆抗体，基于流式细胞术和基于ELISA的诊断平台的检测限分别为0.1和1 ng/mL。所收集的临床患者血清样本也通过这两种血清分型诊断平台进行检测。检测登革病毒1型NS1蛋白的灵敏度和特异性分别为90%和96%。我们的平台检测临床样本的准确性比两种商业NS1诊断平台更先进。总之，我们的平台适用于早期检测登革病毒1型感染患者中的NS1蛋白。
