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Quantum Dot Nanobeads-Labelled Lateral Flow Immunoassay Strip for Rapid and Sensitive Detection of Salmonella Typhimurium Based on Strand Displacement Loop-Mediated Isothermal Amplification
Engineering ( IF 12.8 ) Pub Date : 2021-07-17 , DOI: 10.1016/j.eng.2021.03.024
Yuting Shang 1, 2 , Shuzhen Cai 1 , Qinghua Ye 1 , Qingping Wu 1 , Yanna Shao 1 , Xiaoying Qu 1 , Xinran Xiang 1 , Baoqing Zhou 1 , Yu Ding 1 , Moutong Chen 1 , Liang Xue 1 , Honghui Zhu 1 , Jumei Zhang 1
Affiliation  

Rapid, sensitive, point-of-care detection of pathogenic bacteria is important for food safety. In this study, we developed a novel quantum dot nanobeads-labelled lateral flow immunoassay strip (QBs-labelled LFIAS) combined with strand displacement loop-mediated isothermal amplification (SD-LAMP) for quantitative Salmonella Typhimurium (ST) detection. Quantum dot nanobeads (QBs) served as fluorescence reporters, providing good detection efficiency. The customizable strand displacement (SD) probe was used in LAMP to improve the specificity of the method and prevent by-product capture. Detection was based on a sandwich immunoassay. A fluorescence strip reader measured the fluorescence intensity (FI) of the test (T) line and control (C) line. The linear detection range of the strip was 102–108 colony forming units (CFU)·mL−1. The visual limit of detection was 103 CFU·mL−1, indicating that the system was ten-fold more sensitive than AuNPs-labelled test strips. ST specificity was analyzed in accordance with agarose gel outputs of polymerase chain reaction (PCR) and SD-LAMP. We detected ST in foods with an acceptable recovery of 85%–110%. The method is rapid, simple, almost equipment-free, and suitable for bacterial detection in foods and for clinical diagnosis.



中文翻译:

基于链位移环介导等温扩增的量子点纳米珠标记侧流免疫检测条用于鼠伤寒沙门氏菌的快速灵敏检测

病原菌的快速、灵敏、床旁检测对于食品安全非常重要。在这项研究中,我们开发了一种新型量子点纳米珠标记的横向流动免疫测定条(QBs 标记的 LFIAS)结合链置换环介导的等温扩增(SD-LAMP)用于定量鼠伤寒沙门氏菌(ST)检测。量子点纳米珠 (QB) 作为荧光报告基因,提供了良好的检测效率。可定制的链置换 (SD) 探针用于 LAMP 以提高方法的特异性并防止副产物捕获。检测基于夹心免疫测定。荧光条读取器测量测试 (T) 线和对照 (C) 线的荧光强度 (FI)。条带的线性检测范围为10 2–10 8菌落形成单位(CFU)·mL −1。视觉检测限为10 3 CFU·mL -1,表明该系统比AuNPs标记的测试条灵敏十倍。根据聚合酶链反应 (PCR) 和 SD-LAMP 的琼脂糖凝胶输出分析 ST 特异性。我们在食品中检测到 ST,可接受的回收率为 85%–110%。该方法快速、简单、几乎不需要设备,适用于食品中的细菌检测和临床诊断。

更新日期:2021-07-17
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