A multiplex PCR assay with a common primer for the detection of eleven foodborne pathogens

Abstract

Salmonella enterica, Listeria monocytogenes, Shigella flexneri, Escherichia coli O157:H7, Vibrio parahaemolyticus, Staphylococcus aureus, Vibrio cholerae, Clostridium botulinum type A, Bacillus cereus, Clostridium perfringens Alpha toxin, and Yersinia enterocolitica are 11 common foodborne pathogens. Traditional bacterial culture methods for detecting pathogens are time‐consuming and labor‐intensive. Multiplex PCR technology, which can detect multiple targets in a single tube, has been increasingly applied to microbial detection due to its high specificity, sensitivity, and fast response. This paper is to establish a multiplex PCR technology mediated by a common primer for the detection of these 11 common foodborne pathogens in order to achieve the goal of nondirectional screening for these 11 common foodborne pathogens. The specificity of the established CP‐MPCR detection system was first verified by 100 clinical isolates. The sensitivity of the CP‐MPCR detection system was then detected by using cultured bacteria preparations and has been confirmed with a high sensitivity of 103 to 104 CFU/mL, among them, the sensitivity of the CP‐MPCR for Vibrio cholerae and S. flexneri can even achieve 102 CFU/mL. Sixty anal swab samples collected from Suzhou CDC and 16 enrichment cultured solutions of food samples collected from the Suzhou Food Inspection and Testing Center were tested using the CP‐MPCR system. A total of 32 positive results were detected. Practical Application Food poisoning incidents occur frequently around the world, mainly because of the contamination of food by pathogenic bacteria and serious harm to human health. The method provided in this study can detect 11 foodborne pathogens in food, which can effectively prevent the spread of pathogenic microorganisms. At the same time, for the food poisoning incident that has already occurred, this method can be used for diagnosis to find out the cause.

CRISPR‐Cas‐based detection for food safety problems: Current status, challenges, and opportunities

“…Foodborne disease is one of the major threats to food safety (Tao et al., 2020); it poses a series of challenges and is a global burden because of its contribution to morbidity and mortality (Zheng et al., 2021). Considering the severity of foodborne diseases caused by pathogens or their toxins, it is imperative to develop novel diagnostic models with specificity and sensitivity (Zheng et al., 2021).…”

Aptasensors for Staphylococcus aureus Risk Assessment in Food

“…With the development of molecular detection technology, PCR has been gradually applied to foodborne pathogenic bacteria detection. However, owing to its inability to distinguish between dead and alive bacteria as well as the fact that the experiment is prone to interference, it is difficult to satisfy the requirements of actual detection ( Tao et al, 2020 ). Compared with traditional antibodies, aptamers, which act as a new molecular recognition element, have the advantages of a wide range of target molecules, fabulous stability, long storage life, and high specificity ( Gopinath et al, 2016 ).…”

“…Multiplex PCR provides faster detection by simultaneously amplifying multiple gene targets compared to simple PCR. Tao et al (2020) established a common primer-mediated MPCR technique for non-qualitative screening of 11 common foodborne pathogens. Park et al (2006) used MPCR to detect S. aureus in kimchi, and the detection limit was 260 CFU/ml.…”

Quantum Dots for Pathogenic Bacterial Monitoring and Combating

“…So more rapid and accurate detecting technologies have been invented. One of the most evolutionary methods is the polymerase chain reaction, [ 47–49 ] which can identify the bacterial classification and provide gene expression level in a few hours with better accuracy. [ 47,50 ] Besides, the coupling of chromatography and mass spectra like matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry [ 51–55 ] can play a more exact role in diagnosis.…”