Development of IMBs-qPCR method for detection of foodborne Salmonella


Citation

Sun, R. and Cao, H. and Fu, Y. and Tan, Z. and Liu, N. and Li, H. and Feng, Z. and Chi, H. and Hua, D. and Huang, J. (2023) Development of IMBs-qPCR method for detection of foodborne Salmonella. International Food Research Journal (Malaysia), 30. pp. 978-991. ISSN 2231 7546

Abstract

Rapid and accurate detection of pathogenic microorganism is critical for food safety. Salmonella is one of the common causes of food poisoning. In the present work, polyclonal antibody against the recombinant PagN protein was prepared, and coupled with carboxylated magnetic beads to form immunomagnetic beads (IMBs) for capturing Salmonella, which was then combined with qPCR technology which used the specific primers of invA gene to accurately quantify the number of colonies, thus establishing the IMBs-qPCR method for detection of Salmonella. 0.2 mg IMBs could specifically concentrate Salmonella, with the stable capturing efficiency of 80%, corresponding to the concentrations of 102 - 105 CFU/mL. The minimum detection limit concentration was 101 CFU/mL. The method was applied for detection and enumeration of Salmonella in pork and milk samples, and the capture efficiency of 77.38 and 80.92% were obtained. In summary, the IMBs-qPCR method established herein could effectively detect Salmonella with good specificity and sensitivity. The whole detection time was less than 9h, which laid a foundation for development of a rapid detection kit for foodborne pathogens.


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Abstract

Rapid and accurate detection of pathogenic microorganism is critical for food safety. Salmonella is one of the common causes of food poisoning. In the present work, polyclonal antibody against the recombinant PagN protein was prepared, and coupled with carboxylated magnetic beads to form immunomagnetic beads (IMBs) for capturing Salmonella, which was then combined with qPCR technology which used the specific primers of invA gene to accurately quantify the number of colonies, thus establishing the IMBs-qPCR method for detection of Salmonella. 0.2 mg IMBs could specifically concentrate Salmonella, with the stable capturing efficiency of 80%, corresponding to the concentrations of 102 - 105 CFU/mL. The minimum detection limit concentration was 101 CFU/mL. The method was applied for detection and enumeration of Salmonella in pork and milk samples, and the capture efficiency of 77.38 and 80.92% were obtained. In summary, the IMBs-qPCR method established herein could effectively detect Salmonella with good specificity and sensitivity. The whole detection time was less than 9h, which laid a foundation for development of a rapid detection kit for foodborne pathogens.

Additional Metadata

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Item Type: Article
AGROVOC Term: Salmonella
AGROVOC Term: pathogens
AGROVOC Term: cultural methods
AGROVOC Term: quantitative polymerase chain reaction
AGROVOC Term: biological contamination
AGROVOC Term: experimental design
AGROVOC Term: sampling
AGROVOC Term: microbiologists
AGROVOC Term: food safety
AGROVOC Term: food enrichment
Geographical Term: China
Depositing User: Nor Hasnita Abdul Samat
Date Deposited: 11 Nov 2025 08:21
Last Modified: 11 Nov 2025 08:21
URI: http://webagris.upm.edu.my/id/eprint/1407

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