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Rapid detection of Mycoplasma pneumoniae by loop-mediated isothermal amplification assay

Xin Yuan1, Changqing Bai, Qian Cui

  • 1Department of Respiratory and Critical Care Medicine, 307th Hospital of Chinese People's Liberation Army Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing City, China.

Medicine
|June 21, 2018
PubMed

Insights

A new loop-mediated isothermal amplification (LAMP) assay rapidly detects Mycoplasma pneumoniae (Mp) infection. This sensitive and specific method offers a viable alternative to quantitative real-time polymerase chain reaction (qRT-PCR) for timely diagnosis.

Area of Science:

  • Microbiology
  • Molecular Diagnostics
  • Infectious Diseases

Background:

  • Mycoplasma pneumoniae (Mp) infections require accurate and rapid diagnostic methods.
  • Antibiotic misuse is a concern in Mp treatment, highlighting the need for precise diagnostics.

Purpose of the Study:

  • To develop a loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Mycoplasma pneumoniae.
  • To evaluate the sensitivity, specificity, and clinical applicability of the developed LAMP assay.

Main Methods:

  • Designed six primers targeting the Mp P1 gene for LAMP assay development.
  • Determined assay sensitivity using serial dilutions of a standard Mp strain.
  • Assessed specificity against 17 common respiratory pathogens.
  • Compared LAMP results with quantitative real-time polymerase chain reaction (qRT-PCR) using 125 clinical samples.

Main Results:

  • The LAMP assay detected Mp within 60 minutes with a minimum detection limit of 39 pg/μL.
  • No cross-reactivity was observed with 17 common respiratory pathogens, indicating high specificity.
  • LAMP assay identified 43 positive cases out of 125 clinical specimens, closely aligning with qRT-PCR results (40 positive cases).
  • Sequencing confirmed Mp in all 43 samples positive by LAMP.

Conclusions:

  • The developed LAMP assay is suitable for the rapid, sensitive, and specific detection of Mycoplasma pneumoniae.
  • This LAMP assay demonstrates comparable performance to qRT-PCR, offering a valuable diagnostic tool.
  • The assay can aid in improving Mp diagnosis and reducing unnecessary antibiotic use.

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