THE APPLICATION OF THE RAPD-PCR METHOD FOR IDENTIFICATION OF LACTIC ACID BACTERIA
І. M. Slyvka1, O. Y. Tsisaryk1, T. Bocer2
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1Lviv National University of Veterinary Medicine and Biotechnologies named
after S. Z. Gzhytskyj, Pekarska st., 50, Lviv, 79010, Ukraine
2University of Rzeszow, Sokolowska st., 26, Kolbuszowa, 36-100, Poland
The heterogeneous properties of lactic acid bacteria (LAB) create a significant difficulties in their identification. The use of genotypic methods based on polymerase chain reaction (PCR) has opened new possibilities for precise and rapid identification of LAB. Our purpose was to identify LAB isolated from ewe’s cheese that is traditionally produced in the Carpathian region of Ukraine. A number of microbiological and molecular genetic studies, such as cultural and morphological properties of the isolates LAB and the method of analysis random amplification of polymorphic DNA (RAPD-PCR) have been used to achieve this purpose. The material for researches was ewe’s cheese buts (brine cheese before ripening) produced in the alpine mountain valley of Putyla district of Chernivtsi region. A pure cultures LAB, which were isolated from the cheese, served as subject of the research.
The reference strains of LAB we used for the comparative analysis of amplified fragments of DNA of the investigated strains of LAB. The investigated strains of LAB were taken to family Lactobacteriaceae and Streptococcaceae under the nature of growth of colonies and the temperature conditions of cultivating and to the genus Lactobacterium, Lactococcus and Streptococcus under the morphological properties of isolates.
The considerable differences in electrophoretic movable were found between the investigated strains of LAB that consisted in the absence of similar, general fragments of DNA, and also a small number of identical DNA fragments were found in some spectra. The absence of an identical DNA profiles indicates heterogeneity properties of isolated LAB and their intraspecific diversity.
RAPD-PCR method is appropriate to identify generic affinity isolates of LAB detection and intraspecific changes but not sufficient for species and strains identification of LAB.
Keywords: LACTIC ACID BACTERIA, GENOTYPING, POLYMERASE CHAIN REACTION, AMPLIFICATION, POLYMORPHISM
