A Rapid PCR Based Method to Distinguish between Lactococcus and Enterococcus

Phenotypic characterisation of Lactococcus and Enterococcus species remains unreliable as strains of both genera have been isolated which do not conform to the traditional criteria for separation of these genera. A bank of 131 isolates was phenotypically characterised by three methods: (a) tradition...

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Bibliographic Details
Published in:Systematic and applied microbiology 2000-12, Vol.23 (4), p.510-522
Main Authors: Deasy, B.M., Rea, M.C., Fitzgerald, G.F., Cogan, T.M., Beresford, T.P.
Format: Article
Language:English
Subjects:
16S rDNA
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biological and medical sciences
DNA Primers - genetics
DNA, Bacterial - genetics
Enterococcus
Enterococcus - classification
Enterococcus - genetics
Food Microbiology
Fundamental and applied biological sciences. Psychology
Genes, Bacterial
Genotype
genus-specific primers
Lactobacillus - classification
Lactobacillus - genetics
Lactococcus
Microbiology
Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains
Phenotype
Phylogeny
Polymerase Chain Reaction - methods
Reagent Kits, Diagnostic
RNA, Ribosomal, 16S
Species Specificity
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Summary:Phenotypic characterisation of Lactococcus and Enterococcus species remains unreliable as strains of both genera have been isolated which do not conform to the traditional criteria for separation of these genera. A bank of 131 isolates was phenotypically characterised by three methods: (a) traditional broth tests, (b) API Rapid ID 32 Strep and (c) BBL Crystal ID kits. Differences in genus designation between commercial kits were evident for 12 strains (9%), while 7 strains (5%) remained unidentified by either kit. Published 16S rRNA sequences were aligned and used to design genus-specific primers which, when used in separate PCR reactions, were capable of distinguishing all type strains of Lactococcus and Enterococcus. These primers did not react with known species of Streptococcus, Pediococcus, Lactobacillus, Leuconostoc or Tetragenococcus. Isolates which could not be identified by phenotype were assigned to either genus on the basis of the gene primers.
ISSN:0723-2020
1618-0984
DOI:10.1016/S0723-2020(00)80025-9