getpdf  NLM-PubMed-Logo  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. Kurbakov1
*orcid tiny, Evgenii A. Konorov1,2,3orcid tiny, Mikhail Y. Minaev1orcid tiny and Oksana A. Kuznetsova1orcid tiny


1V.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
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Key words:
real-time PCR, HRM, starter cultures, fermented sausages, Lactobacillus sakeiLactobacillus 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: tel3 +79629529223
                                          email3 homo_ludens@vniimp.ru

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