Development of IMBs-qPCR detection method for Yersinia enterocolitica based on the foxA gene.

Jingxuan Shi, Heng Chi, Aiping Cao, Yinna Song, Min Zhu, Lilin Zhang, Fuzhou Xu, Jinhai Huang

Archives of microbiology 2021 Sep

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Yersinia enterocolitica is an important zoonotic pathogen, which seriously endangers food-safety risk. In this study, the recombinant outer membrane protein OmpF and its antibody were prepared and coupled with immunomagnetic beads (IMBs) to capture Y. enterocolitica in food samples, combining the quantitative PCR detection with primers of virulence factor gene foxA for Yersinia enterocolitica contamination. The results showed that the capture efficiency of approximately 80% using anti-OmpF antibody-immunomagnetic beads and linearly dependent capture under 101-105 CFU/mL Y. enterocolitica compared with less than 10% capture of other bacteria. The detection limit of 64 CFU/mL was obtained based on foxA gene PCR detection combined with capture of the anti-OmpF antibody-immunomagnetic beads to detect Yersinia enterocolitica in artificially contaminated milk and pork samples. Compared to the culture method, the developed IMBs-qPCR method has higher consistency, was less time consuming, which taken together provides an effective alternative method for rapid detection of Y. enterocolitica in food. © 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.

Citation

Jingxuan Shi, Heng Chi, Aiping Cao, Yinna Song, Min Zhu, Lilin Zhang, Fuzhou Xu, Jinhai Huang. Development of IMBs-qPCR detection method for Yersinia enterocolitica based on the foxA gene. Archives of microbiology. 2021 Sep;203(7):4653-4662