İnsan bağırsağı kaynaklı lactobacıllus türlerinin pcr-ardra tekniği ile tanımlanması

Probiotic means 'living microorganisms' which have beneficial effects on their host's health. In recent years, interest in the probiotic lactobacilli has been stimulated by the use of these bacteria in products that are claimed to confer health benefits on the consumer. Lactobacillus is among the most important organisms with nearly 140 species at present, mostly of industrial and clinical importance. The identification of probiotic strain only in phenotypic and physiyological characteristics is often with a low level of discrimination. The identification of Lactobacillus at species level is becoming more and more required. Unfortunately, most of the methods are labor-intensive, costly, and time-consuming. The aim of this study is to identify and discriminate fourteen Lactobacillus strains that have been found in the human alimentary tract by the use of amplified 16S ribosomal DNA restriction analysis (ARDRA). In this study, using two universal primers (8AU and 1492R) against the 16S rRNA gene, 1.5 kb PCR products were obtained from the commercial strains of the manufacturers. To obtain species-specific restriction patterns, three restriction endonucleases (FspBI, HinfI and DraI) were chosen for the aligning of the 16S rRNA gene sequences of fourteen Lactobacillus species retrieved from various databases. It was shown that 1.5 kb amplicon digested by these enzymes provided unique patterns of almost fourteen species. Digestion of the 1.5 kb PCR product with FspBI, HinfI and DraI endonucleases differentiated six, four and two species of Lactobacillus respectively. The present study has demonstrated that 1.5 kb 16S rRNA gene fragments can be identified and differentiated in a reliable, rapid and accurate manner for Lactobacillus species that are found in the human alimentary tract.