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Related Experiment Videos

A rapid PCR based method to distinguish between Lactococcus and Enterococcus.

B M Deasy1, M C Rea, G F Fitzgerald

  • 1Dairy Products Research Centre, Fermoy, Co. Cork, Ireland.

Systematic and Applied Microbiology
|March 16, 2001
PubMed
Summary
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Phenotypic identification of Lactococcus and Enterococcus bacteria is unreliable. Genus-specific PCR primers targeting 16S rRNA sequences accurately differentiate these bacteria, improving diagnostic accuracy.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Bacterial Taxonomy

Background:

  • Traditional phenotypic methods for differentiating Lactococcus and Enterococcus species are often unreliable.
  • Some bacterial strains exhibit characteristics that do not conform to established criteria for genus separation.
  • Commercial identification kits show discrepancies and limitations in accurately classifying these bacteria.

Purpose of the Study:

  • To evaluate the reliability of phenotypic characterization for Lactococcus and Enterococcus species.
  • To develop and validate a molecular method for accurate genus-specific identification.
  • To improve the diagnostic accuracy for distinguishing between Lactococcus and Enterococcus.

Main Methods:

  • Phenotypic characterization of 131 bacterial isolates using traditional broth tests and commercial kits (API Rapid ID 32 Strep, BBL Crystal ID).

Related Experiment Videos

  • Design of genus-specific primers based on aligned 16S rRNA gene sequences.
  • Polymerase Chain Reaction (PCR) using designed primers for differentiating Lactococcus and Enterococcus type strains and isolates.

    • Main Results:

    • Phenotypic methods showed inconsistencies, with 9% of strains misidentified and 5% unidentified by commercial kits.
    • Genus-specific PCR primers successfully distinguished all tested Lactococcus and Enterococcus type strains.
    • The developed primers did not cross-react with other related genera (Streptococcus, Pediococcus, Lactobacillus, Leuconostoc, Tetragenococcus).

    Conclusions:

    • Phenotypic characterization is insufficient for reliable differentiation of Lactococcus and Enterococcus.
    • Genus-specific PCR primers targeting 16S rRNA sequences provide a robust and accurate method for distinguishing these genera.
    • Molecular identification using these primers resolves ambiguities encountered with phenotypic methods.