https://doi.org/10.17113/ftb.57.01.19.5983

       

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


Konstantin A. Kurbakov^1*, Evgenii A. Konorov^1,2,3, Mikhail Y. Minaev¹ and Oksana A. Kuznetsova<sup>1


1</sup>V.M. Gorbatov Federal Research Center for Food Systems of Russian Academy of Sciences, Talalikhina 26, 109316, Moscow, Russian Federation
²Vavilov Institute of General Genetics, Gubkina 3, 119333, Moscow, Russian Federation
³Martsinovsky Institute of Medical Parasitology, Tropical and Vector-Borne Diseases, Sechenov University, Malaya Pirogovskaya 20-1, 119435 Moscow, Russian Federation



Article history:
Received: 9 August 2018
Accepted: 17 January 2019




Key words:
real-time PCR, HRM, starter cultures, fermented sausages, Lactobacillus sakei, Lactobacillus curvatus

Summary:
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.

*Corresponding author:  +79629529223
                                           homo_ludens@vniimp.ru