Lateral Flow Assay with Near-Infrared Dye for Multiplex Detection
Lateral Flow Assay with Near-Infrared Dye for Multiplex Detection
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DOI：
10.1373/clinchem.2012.200360
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发表时间：
2013-04-01
影响因子：
9.3
通讯作者：
D'Andrea, Annalisa
中科院分区：
文献类型：
--
作者：
Swanson, Christina;D'Andrea, Annalisa
背景：横向流动测定法（LFAs）是流行的即时诊断工具，因为它们快速且易于使用。然而，它们通常缺乏分析灵敏度和定量输出，并且可能难以复用，限制了它们在生物标志物测量中的有用性。作为一项概念验证研究，我们详细介绍了一种定量的、多路LFA的设计，该设计具有现成的近红外（NIR）检测，以提高分析灵敏度。方法：将近红外染料与选定的抗体偶联，结合到LFAs中。我们采用单路优化的NIR-LFAs检测白细胞介素(IL)-6的范围为0 ~ 200 pg/mL，并开发了双路检测方法，在单个试纸上同时检测IL-6的范围为0 ~ 100 pg/mL （0 ~ 4.5 pmol/L）和c反应蛋白（CRP）的范围为50 ~ 2500 ng/mL （0.4 ~ 20 nmol/L）。对60种不同的加标样品进行了检测，并与ELISA结果进行了比较。结果：NIR-LFAs在10%血浆基质中检测到IL-6，检测限为4 pg/mL (182 fmol/L)， CV为1
BACKGROUND: Lateral flow assays (LFAs) are popular point-of-care diagnostic tools because they are rapid and easy to use. Nevertheless, they often lack analytical sensitivity and quantitative output and may be difficult to multiplex, limiting their usefulness in biomarker measurement. As a proof-of-concept study, we detail the design of a quantitative, multiplex LFA with readily available near-infrared (NIR) detection to improve analytical sensitivity.METHODS: NIR dye was conjugated to selected antibodies and incorporated into LFAs. We used singleplex, optimized NIR-LFAs to measure interleukin (IL)-6 from 0 to 200 pg/mL and developed duplex assays to simultaneously measure IL-6 from 0 to 100 pg/mL (0 to 4.5 pmol/L) and C-reactive protein (CRP) from 50 to 2500 ng/mL (0.4 to 20 nmol/L) on a single test strip. Assays were tested on 60 different spiked samples and compared to ELISA results.RESULTS: NIR-LFAs detected IL-6 in a 10% plasma matrix with a limit of detection of 4 pg/mL (182 fmol/L) and a CV