Identification of Lactobacillus species using tDNA-PCR.
tDNA intergenic spacer PCR (tDNA-PCR) using consensus primers complementary to the conserved edges of the tRNA genes can amplify the intergenic spacers. Separation of the PCR products with capillary electrophoresis enables discrimination between fragments differing only one basepair in length. This method was applied to a collection of 82 Lactobacillus strains belonging to 37 species in order to evaluate the discriminatory power of this technique within this genus. Twenty-one species could be distinguished to species level on the basis of a unique tDNA fingerprint pattern. The other species grouped by two (e.g. L. fermentum and L. cellobiosus) or three (L. acidophilus, L. gallinarum and L. helveticus). Inclusion of the resulting fingerprints in a numerical database containing fingerprints of numerous other Gram-positive and Gram-negative species makes the identification of unknown strains possible.
Baele
Margo
M
Department of Pathology, Bacteriology and Poultry Diseases, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium. Margo.Baele@rug.ac.be
Vaneechoutte
Mario
M
Verhelst
Rita
R
Vancanneyt
Marc
M
Devriese
Luc A
LA
Haesebrouck
Freddy
F
D015373:Bacterial Typing Techniques / Q000379:methods*; D016172:DNA Fingerprinting / Q000592:standards; D004269:DNA, Bacterial / Q000032:analysis*; D021901:DNA, Intergenic / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D016208:Databases, Factual; D019075:Electrophoresis, Capillary / Q000379:methods; D006801:Humans; D007778:Lactobacillus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012343:RNA, Transfer / Q000032:analysis*; D015203:Reproducibility of Results
D004269:DNA, Bacterial; D021901:DNA, Intergenic; D004275:DNA, Ribosomal; D012343:RNA, Transfer