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Related Experiment Videos

Use of PCR and sodium dodecyl sulfate-polyacrylamide gel electrophoresis techniques for differentiation of Prevotella

T Premaraj1, N Kato, K Fukui

  • 1Institute of Anaerobic Bacteriology, Gifu University School of Medicine, 40 Tsukasa-machi, Gifu 500-8705, Japan.

Journal of Clinical Microbiology
|March 13, 1999
PubMed
Summary

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Polymerase chain reaction (PCR) assays effectively differentiate Prevotella intermedia and Prevotella nigrescens. These rapid and reliable PCR methods are suitable for clinical use, unlike the lipase test or SDS-PAGE.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Clinical Diagnostics

Background:

  • Accurate identification of Prevotella intermedia and Prevotella nigrescens is crucial for clinical and research applications.
  • Existing differentiation methods, such as the lipase test and SDS-PAGE, have shown limitations in specificity and reliability.

Purpose of the Study:

  • To develop and evaluate novel polymerase chain reaction (PCR) assays for the specific discrimination of Prevotella intermedia and Prevotella nigrescens.
  • To compare the efficacy of the developed PCR assays against established PCR techniques and phenotypic methods like SDS-PAGE and the lipase test.

Main Methods:

  • Design of species-specific primers based on 16S rRNA sequences for Prevotella intermedia and Prevotella nigrescens.
  • Application of PCR assays using newly designed primers and primers from Guillot and Mouton (1997).

Related Experiment Videos

  • Testing specificity with reference strains, clinical isolates of P. intermedia sensu lato, other Prevotella species, and common oral flora.
  • Comparative analysis with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the lipase test.

    • Main Results:

    • Both developed PCR assays provided unambiguous differentiation between Prevotella intermedia and Prevotella nigrescens, with results consistent with each other.
    • The SDS-PAGE method demonstrated questionable reliability, misidentifying some strains when compared to PCR results.
    • The lipase test yielded positive results for both species, indicating its lack of value in differentiating these two bacteria.

    Conclusions:

    • The newly designed PCR assays are simple, rapid, reliable, and specific for differentiating Prevotella intermedia and Prevotella nigrescens.
    • These PCR methods are suitable for use in clinical studies.
    • The lipase test is not valuable for distinguishing between these two Prevotella species, and the reliability of SDS-PAGE is questionable.