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一种基于侧向流动分析和近红外荧光染料的登革热病毒1快速即时检测方法。

A rapid point-of-care test for dengue virus-1 based on a lateral flow assay with a near-infrared fluorescent dye.

作者信息

Chen Lin, Wang Huagui, Guo Tongsheng, Xiao Chaohui, Liu Licheng, Zhang Xiaoguang, Liu Bo, Li Peiran, Liu Aixia, Li Bo, Li Boan, Mao Yuanli

机构信息

Medical University of PLA, Beijing, China; Center for Clinical Laboratory, 302 Hospital of PLA, Beijing, China.

Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China; Beijing Macro & Micro Test Biotech Co., Ltd, Beijing, China.

出版信息

J Immunol Methods. 2018 May;456:23-27. doi: 10.1016/j.jim.2018.02.005. Epub 2018 Feb 15.

Abstract

Dengue fever is caused by the dengue virus (DENV), and DENV1 is the prevalent epidemic serotype in south China. A new lateral flow assay (LFA) based on a near-infrared (NIR) fluorescent dye was developed to detect anti-DENV1 IgG antibodies. DyLight-800 was used as the marker conjugated to goat anti-human IgG antibodies, and recombinant dengue type 1 envelope protein was used as the capture protein on the test line. Twenty samples from patients infected with DENV1 and 160 negative controls were analyzed using this new NIR-LFA. The results of the NIR-LFA were compared with the results of Panbio Dengue IgG ELISA and the Dengue Duo IgM/IgG Cassette. Nineteen confirmed DENV1-positive samples were identified by NIR-LFA, giving 95% (19/20) sensitivity. No significant differences existed in the results when the 20 primary clinical samples were analyzed using NIR-LFA, Panbio ELISA, and the Dengue Duo Cassette. However, NIR-LFA had a lower limit of detection than IgG ELISA and Duo IgM/IgG Cassette did when analyzing a 2-fold dilution series of the 19 samples positively identified by NIR-LFA. When incorporated with an NIR POCT device, the new NIR-LFA was rapid, easy to use, and highly sensitive in detecting DENV1, and has potential for application to clinical diagnosis.

摘要

登革热由登革病毒(DENV)引起,DENV1是中国南方流行的血清型。开发了一种基于近红外(NIR)荧光染料的新型侧向流动分析法(LFA)来检测抗DENV1 IgG抗体。DyLight-800用作与山羊抗人IgG抗体偶联的标记物,重组登革热1型包膜蛋白用作检测线上的捕获蛋白。使用这种新型NIR-LFA分析了20份感染DENV1患者的样本和160份阴性对照。将NIR-LFA的结果与Panbio登革热IgG ELISA和登革热双联IgM/IgG检测卡的结果进行比较。NIR-LFA鉴定出19份确诊的DENV1阳性样本,灵敏度为95%(19/20)。使用NIR-LFA、Panbio ELISA和登革热双联检测卡分析20份原始临床样本时,结果无显著差异。然而,在分析由NIR-LFA阳性鉴定的19份样本的2倍稀释系列时,NIR-LFA的检测下限低于IgG ELISA和双联IgM/IgG检测卡。与NIR即时检测设备结合使用时,新型NIR-LFA检测DENV1快速、易用且高度灵敏,具有临床诊断应用潜力。

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