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

• Published in: Archives of microbiology Vol. 203; no. 7; pp. 4653 - 4662
• Main Authors: Shi, Jingxuan, Chi, Heng, Cao, Aiping, Song, Yinna, Zhu, Min, Zhang, Lilin, Xu, Fuzhou, Huang, Jinhai
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.09.2021; Springer Nature B.V
• Subjects: Antibodies; Beads; Biochemistry; Biomedical and Life Sciences; Biotechnology; Cell Biology; Ecology; Food; Food contamination; Food contamination & poisoning; Food safety; FoxA gene; Life Sciences; Membrane proteins; Microbial Ecology; Microbiology; Original Paper; Pork; Virulence; Virulence factors; Yersinia enterocolitica; Quantitative real-time PCR; Immunomagnetic beads; Outer membrane protein
• ISSN: 0302-8933; 1432-072X
• DOI: 10.1007/s00203-021-02459-4

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 fox A 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 10 1 –10 5  CFU/mL Y. enterocolitica compared with less than 10% capture of other bacteria. The detection limit of 64 CFU/mL was obtained based on fox A 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.