High-resolution melting real-time polymerase chain reaction assays for subtyping of five diarrheagenic Escherichia coli by a single well in milk.

Autor:	Shan S; College of Life Science, National R&D Center for Freshwater Fish Processing, Jiangxi Normal University, Nanchang 330022, China; Jiangxi Province Key Laboratory of Diagnosing and Tracing of Foodborne Disease, Jiangxi Provincial Center for Disease Control and Prevention, Nanchang 330029, China., Li R; Jiangxi Province Key Laboratory of Diagnosing and Tracing of Foodborne Disease, Jiangxi Provincial Center for Disease Control and Prevention, Nanchang 330029, China; Jiangxi Provincial Key Laboratory of Preventive Medicine, School of Public Health, Nanchang University, Nanchang 330019, China., Xia W; Center for Life Sciences, Yunnan University, Kunming 650031, China., Tong X; College of Life Science, National R&D Center for Freshwater Fish Processing, Jiangxi Normal University, Nanchang 330022, China., Huang Y; Jiangxi YeLi Medical Device Co. Ltd., Nanchang 330008, China., Tan Y; Jiangxi Province Key Laboratory of Diagnosing and Tracing of Foodborne Disease, Jiangxi Provincial Center for Disease Control and Prevention, Nanchang 330029, China; Jiangxi Provincial Key Laboratory of Preventive Medicine, School of Public Health, Nanchang University, Nanchang 330019, China., Peng S; Jiangxi Province Key Laboratory of Diagnosing and Tracing of Foodborne Disease, Jiangxi Provincial Center for Disease Control and Prevention, Nanchang 330029, China., Liu C; Jiangxi Province Key Laboratory of Diagnosing and Tracing of Foodborne Disease, Jiangxi Provincial Center for Disease Control and Prevention, Nanchang 330029, China., Wang S; Jiangxi Key Laboratory for Mass Spectrometry and Instrumentation, East China University of Technology, Nanchang 330013, China., Liu D; Jiangxi Province Key Laboratory of Diagnosing and Tracing of Foodborne Disease, Jiangxi Provincial Center for Disease Control and Prevention, Nanchang 330029, China. Electronic address: defoelau@163.com.
Jazyk:	angličtina
Zdroj:	Journal of dairy science [J Dairy Sci] 2024 Aug; Vol. 107 (8), pp. 5416-5426. Date of Electronic Publication: 2024 Mar 14.
DOI:	10.3168/jds.2024-24331
Abstrakt:	Diarrheagenic Escherichia coli (DEC) is a kind of foodborne pathogen that poses a significant threat to both food safety and human health. To address the current challenges of high prevalence and difficult subtyping of DEC, this study developed a method that combined multiplex PCR with high-resolution melting (HRM) analysis for subtyping 5 kinds of DEC. The target genes are amplified by multiplex PCR in a single well, and HRM curve analysis was applied for distinct amplicons based on different melting temperature (Tm) values. The method enables discrimination of different DEC types based on characteristic peaks and distinct Tm values in the thermal melting curve. The assay exhibited 100% sensitivity and 100% specificity with a detection limit of 0.5 to 1 ng/μL. The results showed that different DNA concentrations did not influence the subtyping results, demonstrating this method owed high reliability and stability. In addition, the method was also used for the detection and subtyping of DEC in milk. This method streamlines operational procedures, shorts the detection time, and offers a novel tool for subtyping DEC. (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)
Databáze:	MEDLINE
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