Rapid identification of dairy lactic acid bacteria by M13-generated, RAPD-PCR fingerprint databases.
About a thousand lactic acid bacteria (LAB) isolated from dairy products, especially cheeses, were identified and typed by species-specific PCR and RAPD-PCR, respectively. RAPD-PCR profiles, which were obtained by using the M13 sequence as a primer, allowed us to implement a large database of different fingerprints, which were analysed by BioNumerics software. Cluster analysis of the combined RAPD-PCR fingerprinting profiles enabled us to implement a library, which is a collection of library units, which in turn is a selection of representative database entries. A library unit, in this case, can be considered to be a definable taxon. The strains belonged to 11 main RAPD-PCR fingerprinting library units identified as Lactobacillus casei/paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus helveticus, Lactobacillus delbrueckii, Lactobacillus fermentum, Lactobacillus brevis, Enterococcus faecium, Enterococcus faecalis, Streptococcus thermophilus and Lactococcus lactis. The possibility to routinely identify newly typed, bacterial isolates by consulting the library of the software was valued. The proposed method could be suggested to refine previous strain identifications, eliminate redundancy and dispose of a technologically useful LAB strain collection. The same approach could also be applied to identify LAB strains isolated from other food ecosystems.
Rossetti
Lia
L
Istituto Sperimentale Lattiero Caseario, Via Lombardo 11, 26900 Lodi, Italy.
Giraffa
Giorgio
G
D000818:Animals; D015373:Bacterial Typing Techniques; D017104:Bacteriophage M13 / Q000235:genetics*; D002611:Cheese / Q000382:microbiology*; D016172:DNA Fingerprinting / Q000379:methods; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D015723:Gene Library; D007778:Lactobacillus / Q000145:classification* / Q000235:genetics / Q000821:virology; D008892:Milk / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods*; D017422:Sequence Analysis, DNA; D012984:Software; D013045:Species Specificity; D013289:Streptococcaceae / Q000145:classification* / Q000235:genetics / Q000821:virology; D013997:Time Factors
D017931:DNA Primers; D004269:DNA, Bacterial