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基于杂交链式反应的适体传感器用于脂多糖的高灵敏检测。

Highly sensitive detection of lipopolysaccharides using an aptasensor based on hybridization chain reaction.

机构信息

Beijing Laboratory for Food Quality and Safety, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.

Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science &Nutritional Engineering, China Agricultural University, Beijing, 100083, China.

出版信息

Sci Rep. 2016 Jul 12;6:29524. doi: 10.1038/srep29524.

Abstract

Lipopolysaccharides (LPS), integral components of the outer membrane of all gram-negative bacteria, are closely associated with foodborne diseases such as fever, diarrhea and hypotension, and thus, the early and sensitive detection of LPS is necessary. In this study, an aptasensor assay based on hybridization chain reaction (HCR) was developed to detect LPS. Briefly, two complementary stable species of biotinylated DNA hairpins coexisted in solution until the introduction of a detection probe triggered a hybridization chain reaction cascade. The DNA conjugates specifically reacted with the LPS, which were captured by the ethanolamine aptamer attached to the reaction well surface. After optimizing the key reaction conditions, such as the reaction time of HCR, the amount of the capture probe and detection probes, the increase in the LPS concentration was readily measured by the optical density value, and a relatively low detection limit (1.73 ng/mL) was obtained, with a linear response range of 1-10(5 )ng/mL. The approach presented herein introduced the use of an aptasensor for LPS discrimination and HCR for signal amplification, offering a promising option for detecting LPS.

摘要

脂多糖(LPS)是所有革兰氏阴性菌外膜的组成部分,与食源性疾病如发热、腹泻和低血压密切相关,因此,早期和敏感地检测 LPS 是必要的。在本研究中,开发了一种基于杂交链式反应(HCR)的适体传感器测定法来检测 LPS。简而言之,两种互补的生物素化 DNA 发夹的稳定物种在溶液中共存,直到引入检测探针触发杂交链式反应级联。DNA 缀合物特异性地与 LPS 反应,被附着在反应孔表面上的乙醇胺适体捕获。在优化关键反应条件(如 HCR 的反应时间、捕获探针和检测探针的量)后,可以通过光密度值轻松测量 LPS 浓度的增加,并且获得相对较低的检测限(1.73ng/mL),线性响应范围为 1-10(5)ng/mL。本研究方法引入了适体传感器用于 LPS 鉴别和 HCR 用于信号放大,为检测 LPS 提供了一种很有前途的选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d89/4941573/7cd1d373f9a6/srep29524-f1.jpg

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