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

Abstract

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 e ciency of approximately 80% using anti-OmpF antibody-immunomagnetic beads and linearly dependent capture under 10 1 -10 5 CFU/mL Y. enterocolitica . compare with less than 10% capture of other bacteria. The detection limit of 64 CFU/mL was obtained by based on fox A gene PCR detection combined with capture of the anti-OmpF antibodyimmunomagnetic beads to detect Yersinia enterocolitica in arti cially contaminated milk and pork samples. Comparing with the culture method, the developed IMBs-qPCR method has higher consistency, less time consuming, which providing an effective alternative method for rapid detection of Y.

Efficient Hapten-Specific Biopanning Strategy Based on the Fe3O4@ENR-Functionalized Core–Shell Magnetic Nanoparticles

“…In immunomagnetic separation technology, specific crystal forms or oxides of transition metals, such as Fe, Co, and Ni, were usually used as magnetic cores. The covalent binding of antibodies, oligonucleotide chains, or phages was achieved by modifying active groups such as –NH, –OH, –COOH, or other functional groups on the surface of magnetic cores. , The magnetic nanoparticles (MNPs) with specific ligands formed an “immunomagnetic microsphere-target” complex with the target substance through the antigen–antibody specific reaction and moved directionally under the action of a specific magnetic field to achieve the separation of the target substance from the complex. − In recent years, functionalized core–shell magnetic nanoparticles (FMNPs) have developed rapidly in the fields of immunoassay, cell separation, protein purification, etc. In Liu’s research, FMNPs with amino acid using ultrasonic, long-time mixing, and “one-step” approaches were constructed to be utilized for oleaginous microalgae harvesting .…”