关键词: 环介导间接PCR, 肉制品, 单增李斯特菌

Abstract: Objective: To develop a loop-mediated indirect PCR method for rapid detection of L.monocytogene in meat products. Methods: Chelex-100 resin was used to directly extract template DNAs from meat products. Two probes targeted the conserved region of hly A gene of L.monocytogene were designed according to the published sequences and used to label the Lectin gene fragment of soybean (reporter gene). After hybridization with target genes, gap filling and cyclization, the reporter gene was amplified by PCR for detecting the target genes. Results: The developed detection system showed a detection limit of lower than 100 CFU/g (mL) and had no obvious cross-reactivity with other foodborne pathogenic bacteria. The positive rate of 200 meat product samples was detected to be 2.5%, which was consistent with the result obtained by traditional bacterial separation. Conclusion: The loop-mediated indirect PCR method allows the rapid, sensitive and specific detection of L.monocytogene contamination in meat products.

Key words: loop-mediated indirect PCR, meat products, L. monocytogene