Novel cost-efficient real-time PCR assays for detection and quantitation of Listeria monocytogenes

作者：Traunsek Urban^*; Toplak Natasa; Jersek Barbara; Lapanje Ales; Majstorovic Tamara; Kovac Minka

来源：Journal of Microbiological Methods, 2011, 85(1): 40-46.

DOI：10.1016/j.mimet.2011.01.018

• Detection
• Cost-efficient
• Listeria monocytogenes
• Quantitation
• Real-time PCR

Listeriosis is a serious food-borne infection with mortality rates approaching 30%. Therefore, the rapid, costeffective, and automated detection of Listeria monocytogenes throughout the food chain continues to be a major concern. Here we describe three novel quantitative real-time PCR assays for L monocytogenes based on amplification of a target hlyA gene with SYBR Green I chemistry and hydrolysis probe (TaqMan MGB probe). In order to offer sensitive, rapid and robust tool of additional economical value the real-time PCR assays were designed and optimized to only 5 mu l-reactions. All assays were evaluated by using different non-reference Listeria strains isolated from various food matrices. Results demonstrated specificity to L monocytogenes with accurate quantification over a dynamic range of 5-6 log units with R-2 higher than 0.98 and amplification efficiencies reaching above 92%. The detection and quantification limits were as low as 165 genome equivalents. Comparison of novel assays to commercially available TaqMan (R) Listeria monocytogenes Detection Kit and previously published studies revealed similar specificity, sensitivit

• 出版日期2011-4