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可视化等温重组酶聚合酶扩增检测法快速检测阴沟肠杆菌。

Rapid Detection of Enterobacter cloacae With a Visualized Isothermal Recombinase Polymerase Amplification Assay.

机构信息

Department of Medicine Laboratory & Department of Oncology, The Second People's Hospital of Lianyungang (Cancer Hospital of Lianyungang), Lianyungang, China.

Department of Medicine Laboratory, Lianyungang Second People's Hospital, Kangda College of Nanjing Medical University, Lianyungang, China.

出版信息

Curr Microbiol. 2023 Jun 5;80(7):233. doi: 10.1007/s00284-023-03269-1.

Abstract

Enterobacter cloacae exhibits strong adhesion and invasion properties that contribute its ability to infect the host; it is considered an important opportunistic pathogen throughout the world. To control the spread of E. cloacae, simple, rapid, and accurate detection methods are required. Current methods suffer from various shortcomings and do not meet the demand for on-site quickly detection. Using recombinase polymerase amplification combined with lateral flow strip (RPA-LFS), an isothermal detection method was developed to target the outer membrane protein X (ompX) gene of E. cloacae. This reaction can be performed in 30 min at 37 °C. Limit of detection of 10 CFU/reaction was equivalent to that of the qPCR method. The detection accuracy of clinical samples was also equal to that of the qPCR method. In this study, we developed the RPA-LFS assay, which is simple, rapid, accurate, and does not require a laboratory facility. This assay may prove useful for detecting E. cloacae on-site.

摘要

阴沟肠杆菌具有较强的粘附和侵袭特性,使其能够感染宿主,被认为是全球重要的机会致病菌。为了控制阴沟肠杆菌的传播,需要简单、快速、准确的检测方法。目前的方法存在各种缺陷,不能满足现场快速检测的需求。本研究利用重组酶聚合酶扩增结合侧向流条带(RPA-LFS)技术,针对阴沟肠杆菌外膜蛋白 X(ompX)基因建立了一种等温检测方法。该反应可在 37°C 下 30 分钟内完成。检测限为 10 CFU/反应,与 qPCR 法相当。对临床样本的检测准确性也与 qPCR 法相当。本研究建立的 RPA-LFS 检测方法简单、快速、准确,且不需要实验室设备。该方法有望用于现场检测阴沟肠杆菌。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac2c/10240108/d482a6306248/284_2023_3269_Fig1_HTML.jpg

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