Simultaneous detection of Listeria species isolated from meat processed foods using multiplex PCR

Listeriosis is a foodborne disease caused by the pathogenic Listeria monocytogenes and is considered as a serious health problem due to the severity of symptoms and its high mortality rate. Listeria genus is divided into six species and especially L. monocytogenes is an important foodborne pathogen in humans and livestock. Recently, other Listeria species are reported as pathogenic strains in decayed foods and environments as well. High mortality rate of listeriosis demands for rapid methods to detect the potential presence of the food pathogens in the food industry. We have developed a multiplex PCR for rapid and simultaneous detection of six Listeria species including Listeria grayi, Listeria innocua, Listeria ivanovii, L. monocytogenes, Listeria seeligeri and Listeria welshimeri to identify specific Listeria species in processed foods. The optimized multiplex PCR in this study utilized one Listeria genus specific and each Listeria species-specific primer pairs. Each primer pair yields the products of 370-bp for Listeria genus-specific, 201-bp for L. grayi-specific, 749-bp for L. innocua-specific, 463-bp for L. ivanovii-specific, 509-bp for L. monocytogenes-specific, 673-bp for L. seeligeri-specific and 281-bp for L. welshimeri-specific. We have successfully applied multiplex PCR strategy to 93 Listeria isolates from processed meat products to determine specific Listeria species and out of which 81 strains of L. monocytogenes, 10 strains of L. innocua and 2 strains of L. welshimeri were identified. This established multiplex PCR provides rapid and reliable results and will be useful for the detection of Listeria species in contaminated food products and clinical samples.

► We evaluate novel multiplex PCR assay to discriminate six Listeria species.
► Multiplex PCR assay is highly specific and sensitive for six Listeria species.
► We apply multiplex PCR to 93 Listeria strains isolated from meat processed foods.
► We classify 81 of Listeria monocytogenes, 10 of Listeria innocua and 2 of Listeria welshimeri.
► This is a first study to discriminate six Listeria species using multiplex PCR in one assay.