Applicability of a rapid method based on immunomagnetic capture-fluorescent PCR assay for Campylobacter jejuni.

摘要

In order to develop a rapid method which can check Campylobacter jejuni in foods and water effectively, an immunomagnetic capture-fluorescent PCR (IMC-FPCR) method was established in this paper. The reported method involves isolation of the target pathogen by immunocapture prior to the fluorescent PCR step, therefore the immunomagnetic-beads for Campylobacter were developed, and two groups of primer/probe, which targeted for the species special sequence of flaA gene and hipO gene for C. jejuni were designed. Result indicated that the method permits direct detection of the pathogen without an enrichment step and can be performed in approximately 8h. The assay results are positive for all of the isolates of C. jejuni with a detection limit of approximately 10cfu/ml, are negative for Campylobacter coli and several other bacteria. During the experiment of 300 samples of foods, four of them were positive and the result was the same as traditional biochemical identification. Application of the method to 100 samples of water, one of them was positive but the result was negative with traditional method. Thus, the IMC-FPCR assay provide not only a rapid, sensitive method for quantitative detection of C. jejuni, but also a potential method for detecting of C. jejuni of viable but non-culturable (VBNC) state. All rights reserved, Elsevier.