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Multiplex loop-mediated isothermal amplification (multi-LAMP) assay for rapid detection of mcr-1 to mcr-5 in

Lan-Lan Zhong1,2, Qian Zhou3, Cui-Yan Tan3

  • 1Program in Pathobiology, The Fifth Affiliated Hospital, Zhongshan School of Medicine, Sun Yat-Sen University, Guangdong 519000, People's Republic of China.

Infection and Drug Resistance
|July 17, 2019
PubMed

Insights

A new multiplex loop-mediated isothermal amplification (multi-LAMP) assay rapidly detects colistin resistance genes (mcr-1 to mcr-5). This method offers a sensitive and specific tool for surveillance of antimicrobial resistance in clinical settings.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Infectious Diseases

Background:

  • Plasmid-mediated colistin resistance genes (mcr) are a significant global health threat, particularly in carbapenem-resistant Gram-negative bacterial infections.
  • Effective monitoring of mcr gene spread and epidemiology is crucial, especially in primary care settings.
  • Existing detection methods may not be sufficiently rapid or convenient for widespread clinical surveillance.

Purpose of the Study:

  • To develop and validate a restriction endonuclease-based multiplex loop-mediated isothermal amplification (multi-LAMP) assay.
  • To detect multiple mcr genes (mcr-1 to mcr-5) in colistin-resistant bacteria.
  • To provide a convenient and reliable diagnostic tool for primary care institutions.

Main Methods:

  • Development of a triple-LAMP assay for mcr-1, mcr-3, and mcr-4.
  • Development of a double-LAMP assay for mcr-2 and mcr-5.
  • Determination of assay sensitivity and specificity using electrophoresis and visual detection; validation with clinical samples.

Main Results:

  • The developed LAMP assay demonstrated 10-fold greater sensitivity than PCR.
  • High specificity was observed for the screened primers, with distinct band patterns differentiating specific mcr genes.
  • The assay proved rapid, reliable, and consistent with conventional PCR when applied to clinical samples.

Conclusions:

  • The multi-LAMP assay is a promising tool for the detection of mcr genes.
  • Its implementation can aid in preventing drug-resistant bacterial infections through rapid surveillance in primary care hospitals.
  • This study represents the first application of LAMP for mcr-2 to mcr-5 detection and the first use of multi-LAMP for mcr gene detection.
Keywords:
colistin resistanceenzyme digestionmcr genesmulti-LAMPrapid detection

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