关键词: 食源性致病菌, 多重连接探针扩增技术, 食品检测

Abstract: A method for simultaneous detection of 11 common foodborne pathogenic bacteria in food was established based on the multiplex ligation-dependent probe amplification （MLPA）. By designing and synthesizing specific MLPA probes, hybridization, ligation, and PCR amplification reactions with standard strain DNA after high-temperature denaturation were performed. The size and presence of PCR amplification products were analyzed by capillary electrophoresis. The results showed that using 11 pairs of probes to detect single bacterial nucleic acid samples, each analyte had a single peak and no impurity peaks appeared, indicating that there was no cross influence between the 11 pairs of probes and the specificity of the probes was good;using 11 pairs of probes for simultaneous multiplex detection of mixed pathogenic nucleic acid samples, each analyte could obtain amplification peaks of the same size as expected, and the amplification peaks did not interfere with each other. No target amplification peak was amplified in the blank control, indicating that the system could simultaneously detect multiple foodborne pathogenic bacteria. The detection results of this technology were highly specific, with a minimum detectable contamination level of 1.5×10⁵ CFU/mL. As a supplement to traditional microbiological detection techniques, MLPA technology could be applied for early screening of foodborne pathogenic bacteria and monitoring of food microbiological safety risks.

Key words: foodborne pathogenic bacteria, multiplex ligation-dependent probe amplification, food testing