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Amplification of Proteus mirabilis chromosomal DNA using the polymerase chain reaction

M S Mansy1, A A Fadl, M S Ashour

  • 1Department of Pathobiology, University of Connecticut, Storrs, CT 06269-3089, USA.

Molecular and Cellular Probes
|April 20, 1999
PubMed

Insights

A novel polymerase chain reaction (PCR) method was developed for specific detection of Proteus mirabilis. This highly sensitive assay accurately identifies P. mirabilis DNA, even at minimal concentrations.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Proteus mirabilis is an opportunistic pathogen.
  • Accurate and rapid identification of P. mirabilis is crucial for effective treatment and infection control.

Purpose of the Study:

  • To develop and standardize a specific and sensitive polymerase chain reaction (PCR) assay for the detection of Proteus mirabilis.

Main Methods:

  • Development of P. mirabilis-specific primers (MMKAP 1 and MMKAP 2) based on a 3.5-kilobase pair Hind III fragment DNA clone.
  • Standardization of the PCR protocol.
  • Testing the assay on 18 strains of P. mirabilis and other bacterial species.
  • Determination of the assay's sensitivity using varying amounts of target DNA.

Main Results:

  • A P. mirabilis-specific 3.5-kb DNA product was successfully amplified from 18 P. mirabilis strains.
  • No amplification was observed in other Proteus species or bacterial species, confirming specificity.
  • The assay demonstrated high sensitivity, detecting as little as 10 fg of target DNA.

Conclusions:

  • The developed PCR assay is highly specific and sensitive for the detection of Proteus mirabilis.
  • This molecular method offers a reliable tool for the identification of P. mirabilis in clinical and research settings.

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