Multiplex Real-Time PCR with HRM for Detection of Lactobacillus sakei and Lactobacillus curvatus in Food Samples

• Main Authors: Konstantin A. Kurbakov, Evgenii A. Konorov, Mikhail Y. Minaev, Oksana A. Kuznetsova
• Format: Article
• Language: English
• Published: University of Zagreb 2019-01-01
• Series: Food Technology and Biotechnology
• Subjects: real-time PCR; HRM; starter cultures; fermented sausages; Lactobacillus sakei; Lactobacillus curvatus
• Online Access: http://hrcak.srce.hr/file/319530

Optimization of fermentation processes requires monitoring the species composition of starter cultures and their growth during fermentation. Most starter cultures contain closely related species. Nowadays, high-resolution melting (HRM) analysis is extensively used for multiplex identification of closely related species. In the present paper, we applied real-time polymerase chain reaction (PCR) with HRM analysis for the detection and differentiation of Lactobacillus sakei and L. curvatus. A primer pair was selected for the site of the rpoA gene of Lactobacillus spp. Eleven starter cultures and fifteen fermented sausages with a known bacterial composition were successfully tested using real-time PCR with HRM analysis with the developed primer pair.