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Development and Application of Random DIG-Labelled Probes Based on Total Genomic Lactobacillus DNA
Authors:S. T. Danova  G. Stoyancheva  Y. Manasiev  V. Miteva
Affiliation:(1) Department of Microbial Genetics, Institute of Microbiology, Bulgarian Academy of Sciences, 26, Acad. G. Bontchev St., 1113 Sofia, Bulgaria;(2) Department of Biochemistry and Molecular Biology, Pennsylvania State University, 211 South Frear Bldg., University Park, 16802, PA, USA
Abstract:Summary Nine Lactobacillus-specific and non-isotopically (digoxygenin) labelled probes were developed on the basis of Lactobacillus total chromosomal DNA. Their specificity and applicability for Lactobacillus discrimination was proven by DNA–DNA hybridization to reference strains from the American Type Culture Collection (ATCC). The DNA probes were divided into three groups depending on the ability to hybridize to DNA from the same and/or from a group of related Lactobacillus strains. They were assayed in the species-specific detection of vaginal strains from the genus Lactobacillus. Six DNA probes were successfully applied for characterization of 21 newly isolated vaginal Lactobacilli. The species affliation of some isolates was determined. The developed DNA probes were evaluated for usage as a qualitative hybridization test for detection of Lactobacillus species in mixed cultures, obtained directly from vaginal samples without strain isolation.
Keywords:Dot hybridization  genomic DNA probes  Lactobacillus
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