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Rapid Visual Detection of Methicillin-Resistant Staphylococcus aureus in Human Clinical Samples via Closed LAMP Assay

Noora S A Abusheraida1, Asraa A H AlBaker1, Asmaa S A Aljabri1

  • 1College of Health Science, Qatar University, Doha P.O. Box 2713, Qatar.

Microorganisms
|January 23, 2024
PubMed

Insights

A new loop-mediated isothermal amplification (LAMP) method rapidly detects methicillin-resistant Staphylococcus aureus (MRSA) in clinical samples. This colorimetric assay offers high specificity and sensitivity, crucial for combating antimicrobial resistance.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Clinical Diagnostics

Background:

  • Antimicrobial resistance (AMR), especially from methicillin-resistant Staphylococcus aureus (MRSA), is a growing global health crisis.
  • Existing diagnostic methods for MRSA can be time-consuming, delaying appropriate patient treatment.
  • Rapid and accurate detection of MRSA directly from clinical specimens is essential for effective infection control.

Purpose of the Study:

  • To develop and validate a rapid, closed-tube loop-mediated isothermal amplification (LAMP) method for the direct detection of MRSA.
  • To utilize the hydroxy-naphthol blue (HNB) colorimetric dye assay for visual identification of MRSA.
  • To assess the performance of the LAMP assay against conventional PCR and cefoxitin disk diffusion.

Main Methods:

  • A closed-tube loop-mediated isothermal amplification (LAMP) assay was designed to detect the mecA and spa genes specific to MRSA.
  • Hydroxy-naphthol blue (HNB) dye was incorporated for colorimetric detection, enabling visual results.
  • The assay was tested on 125 preidentified S. aureus isolates and 93 clinical samples, with results compared to conventional PCR and cefoxitin disk diffusion.

Main Results:

  • The LAMP assay demonstrated high diagnostic accuracy, with 100% specificity and 91.23% sensitivity for clinical samples when compared to PCR.
  • For clinical isolates, sensitivity reached 97%, with Cohen Kappa values indicating excellent agreement (0.90-0.926).
  • Compared to cefoxitin disk diffusion, the LAMP assay achieved 100% specificity and sensitivity, with a turnaround time under 1 hour.

Conclusions:

  • The closed-tube LAMP method with HNB dye is a rapid, specific, and sensitive technique for direct MRSA detection from clinical samples.
  • This assay offers a significant improvement over existing methods, providing results in under an hour.
  • The developed LAMP assay is a valuable tool for timely diagnosis and management of MRSA infections, aiding in the fight against antimicrobial resistance.
Keywords:
HNBLAMPMRSAMSSAvisual/colorimetric dye

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