关键词: 食源性细菌, 基因芯片, 多重PCR, 特异性基因, 检测

Abstract: It was to develop a microarray technique coupled with multiplex PCR for the detection and identification 11 food-borne pathogens. The specific gene of each pathogen such as Staphylococcus aureus , Shigellae , Vibrio cholerae and Listeria monocytogenes was chosen as the amplification target. The primers and probes were designed by Premier 5.0 and Oligo 6.0, and then were validated by BLAST. The oligonucleotide probes were immobilized on aldehyde-coated slides, and thirteen pairs of primers were divided into three groups. After the mixed PCR products had hybridization with DNA microarray, the hybridization images were acquired by ScanArray software. 11 food-borne bacteria such as Salmonella typhimurium , Clostridium perfringens and Proteus mirabilis were successfully identified by the array, and the detection limit of this assay was around 10--100 pg. The corresponding hybridization maps of six mixed bacteria DNA samples were obtained, and eighteen clinical isolates of Salmonella spp. were successfully tested. The testing process from PCR can be completed within 3 h. The results indicated that the method can be applied to identify 11 food-borne pathogens rapidly and accurately, and can meet the flux detection demand of multiple food-borne bacteria and has good prospect of clinical application.

Key words: Food-borne pathogens, DNA microarray, Multiplex PCR, Specific gene, Detection