The fluorescent probe-based recombinase-aided amplification for rapid detection of Escherichia coli O157:H7

Dan Mu; Donggen Zhou; Guoyang Xie; Ju Liu; Qin Xiong; Xiaoyan Feng; Hengyi Xu

doi:10.1016/j.mcp.2021.101777

The fluorescent probe-based recombinase-aided amplification for rapid detection of Escherichia coli O157:H7

Mol Cell Probes. 2021 Dec:60:101777. doi: 10.1016/j.mcp.2021.101777. Epub 2021 Nov 1.

Authors

Dan Mu¹, Donggen Zhou², Guoyang Xie³, Ju Liu⁴, Qin Xiong⁵, Xiaoyan Feng⁶, Hengyi Xu⁷

Affiliations

¹ State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047, PR China. Electronic address: 2515828160@qq.com.
² Ningbo International Travel Healthcare Center (Ningbo Customs Port Outpatient Department), Ningbo, 315010, PR China. Electronic address: dongdong1004@163.com.
³ State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047, PR China. Electronic address: 2731024862@qq.com.
⁴ State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047, PR China. Electronic address: 617018708@qq.com.
⁵ State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047, PR China. Electronic address: 1094395604@qq.com.
⁶ State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047, PR China. Electronic address: 1361886215@qq.com.
⁷ State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, 330047, PR China. Electronic address: kidyxu@163.com.

PMID: 34737039
DOI: 10.1016/j.mcp.2021.101777

Abstract

Escherichia coli O157:H7 (E. coli O157:H7) is a common foodborne morbigenous microorganism, which can spread through fecal-oral transmission. Humans can be infected by ingesting foods and water contaminated with E. coli O157:H7, which can cause various symptoms. In present study, we have successfully developed a quick and hypersensitive fluorescent probe-based Recombinase-aided amplification (RAA) method and applied in E. coli O157:H7 detection at 39 °C in 20 min. The sensitivity of the assay in pure E. coli O157:H7 suspension was 5.6 × 10⁰ CFU/mL. The fluorescent probe-based RAA assay was further applied in three samples, and the limit of detection (LOD) in skimmed milk, lettuces and lake water was 5.4 × 10¹ CFU/mL, 7.9 × 10¹ CFU/mL and 5.2 × 10¹ CFU/mL, separately. This method showed a high sensitivity and short detection time, which has the feasible application in on-site test in real samples.

Keywords: Applications; E. coli O157:H7; Isothermal amplification; Recombinase-aided amplification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Escherichia coli O157* / genetics
Fluorescent Dyes
Food Microbiology
Humans
Milk
Recombinases

Substances

Fluorescent Dyes
Recombinases