Development of a simultaneous identification method for nine species of foodborne bacterial pathogens using real-time PCR assays and melting curve analysis

Food safety is a global public health concern. The disease caused by foodborne pathogens often occurs throughout the world and produces considerable economic losses. Hence, rapid detection of these pathogens will enable immediate measures to be implemented for preventing widespread transmission. The...

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Bibliographic Details
Published inJournal of biotech research Vol. 11; pp. 73 - 81
Main Authors Sun, Feilong, Cao, Zhuosong, Qiu, Zhigang, Li, Junwen
Format Journal Article
LanguageEnglish
Published Edmond Bio Tech System 01.01.2020
Subjects
Bacteria
Deoxyribonucleic acid
Design
DNA
DNA polymerase
Economic impact
Environmental management
Food
Food contamination & poisoning
Foodborne diseases
Foodborne pathogens
Identification methods
Melting
Melting curve
Melting point
Melting points
Methods
Pathogens
Polymerase chain reaction
Public health
Real time
Virulence
United States > US
China
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Summary:Food safety is a global public health concern. The disease caused by foodborne pathogens often occurs throughout the world and produces considerable economic losses. Hence, rapid detection of these pathogens will enable immediate measures to be implemented for preventing widespread transmission. The present study was conducted to establish a novel multi-real-time polymerase chain reaction method to detect and quantify nine species of common bacterial pathogens simultaneously from food samples. The nine pairs of primers were designed according to the virulence-associated, high conservative, and specific genes of these pathogens. The products formed were identified based on a melting point temperature (Tm) curve analysis. The food samples with seeded bacterial pathogens were examined. It took about three hours to perform detection. The results confirmed that the assay has good sensitivity and specificity, which can detect the limit to approximately 7x10° copies per reaction respectively. This method would be a routine and practical protocol for identifying and quantifying bacterial pathogens from food or environmental samples and it should facilitate the detection and management of foodborne infectious pathogen outbreaks.
ISSN:1944-3285