Comparison of ribotyping, randomly amplified polymorphic DNA analysis, and pulsed-field gel electrophoresis in typing of Lactobacillus rhamnosus and L. casei strains.
A total of 24 strains, biochemically identified as members of the Lactobacillus casei group, were identified by PCR with species-specific primers. The same set of strains was typed by randomly amplified polymorphic DNA (RAPD) analysis, ribotyping, and pulsed-field gel electrophoresis (PFGE) in order to compare the discriminatory power of the methods. Species-specific primers for L. rhamnosus and L. casei identified the type strain L. rhamnosus ATCC 7469 and the neotype strain L. casei ATCC 334, respectively, but did not give any signal with the recently revived species L. zeae, which contains the type strain ATCC 15820 and the strain ATCC 393, which was previously classified as L. casei. Our results are in accordance with the suggested new classification of the L. casei group. Altogether, 21 of the 24 strains studied were identified with the species-specific primers. In strain typing, PFGE was the most discriminatory method, revealing 17 genotypes for the 24 strains studied. Ribotyping and RAPD analysis yielded 15 and 12 genotypes, respectively.
Tynkkynen
S
S
Valio Ltd. Research and Development Centre, FIN-00039 Valio, Finland. soile.tynkkynen@valio.fi
Satokari
R
R
Saarela
M
M
Mattila-Sandholm
T
T
Saxelin
M
M
D015373:Bacterial Typing Techniques; D017931:DNA Primers; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field; D020459:Genes, rRNA; D005838:Genotype; D007778:Lactobacillus / Q000145:classification* / Q000235:genetics*; D007780:Lacticaseibacillus casei / Q000145:classification / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D019105:Random Amplified Polymorphic DNA Technique; D013045:Species Specificity
D017931:DNA Primers; D015246:Deoxyribonuclease EcoRI