Provincial Key Laboratory for Transfusion-Transmitted Infectious Diseases, Institute of Blood Transfusion, Chinese Academy of Medical Sciences and Peking Union Medical College, Chengdu, Sichuan, China.
Clinical Laboratory Department, Yan'an Hospital Affiliated to Kunming Medical University, Kunming, Yunnan, China.
Int J Infect Dis. 2021 Nov;112:55-62. doi: 10.1016/j.ijid.2021.09.015. Epub 2021 Sep 10.
Shigella flexneri (S. flexneri) is prevalent worldwide and the most common Shigella in many countries, causing highly contagious diarrhea, which seriously threatens public health. This study aimed to develop a colorimetric loop-mediated isothermal amplification (LAMP) for the rapid, accurate, and visualization detection of S. flexneri.
According to the screened specific genes of S. flexneri, three groups of LAMP primers were designed and evaluated, and the colorimetric LAMP reaction volume was optimized. The specificity of the colorimetric LAMP was validated by 20 S. flexneri and 96 non-S. flexneri clinical isolates. In addition, the sensitivity of the developed assay was evaluated by the serial 10-fold dilutions of plasmid DNA.
A colorimetric LAMP assay was developed based on the specific S. flexneri hypothetical protein gene (Accession: AE014073 Region: 4170556.4171068). The colorimetric LAMP method had good specificity for detecting S. flexneri and enabled detection of S. flexneri within 30 minutes, with a plasmid detection limit of 7*10° copies/μL. The results of amplification could be easily identified by color.
This colorimetric LAMP assay could be used for rapid and accurate diagnosis of S. flexneri infection, especially in remote hospitals and laboratories with under-equipped medical facilities, and in situations where an urgent diagnosis is needed.
福氏志贺菌（S. flexneri）在世界范围内广泛流行，是许多国家最常见的志贺氏菌，可引起高度传染性腹泻，严重威胁公众健康。本研究旨在开发一种用于快速、准确和可视化检测 S. flexneri 的比色环介导等温扩增（LAMP）方法。
根据筛选出的 S. flexneri 特异性基因，设计并评估了三组 LAMP 引物，并优化了比色 LAMP 反应体积。通过 20 株 S. flexneri 和 96 株非 S. flexneri 临床分离株验证了比色 LAMP 的特异性。此外，通过质粒 DNA 的连续 10 倍稀释评估了所开发检测方法的灵敏度。
基于 S. flexneri 假定蛋白基因（注册号：AE014073 区域：4170556.4171068），开发了一种比色 LAMP 检测方法。比色 LAMP 方法对检测 S. flexneri 具有良好的特异性，可在 30 分钟内检测到 S. flexneri，质粒检测限为 7*10°拷贝/μL。扩增结果可通过颜色轻松识别。
这种比色 LAMP 检测方法可用于快速准确地诊断 S. flexneri 感染，特别是在医疗设备不足的偏远医院和实验室，以及需要紧急诊断的情况下。
