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Rapid and visual Chlamydia trachomatis detection using loop-mediated isothermal amplification and hydroxynaphthol

I Choopara1, N Arunrut2, W Kiatpathomchai2

  • 1Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, Thailand.

Letters in Applied Microbiology
|October 1, 2016
PubMed
Summary

A new, inexpensive assay using heat-treated DNA and loop-mediated isothermal amplification can rapidly detect Chlamydia trachomatis. This point-of-care test offers comparable accuracy to PCR, making it suitable for resource-limited settings.

Keywords:
Chlamydia trachomatiscolorimetric detectionhydroxynaphthol blueloop-mediated isothermal amplificationsexually transmitted disease

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Area of Science:

  • Molecular Biology
  • Infectious Diseases
  • Diagnostics

Background:

  • Chlamydia trachomatis is a leading cause of bacterial sexually transmitted infections globally.
  • Untreated infections can lead to severe long-term health consequences including infertility and increased cancer risk.
  • Accurate and accessible diagnostic tools are crucial for effective management and control.

Purpose of the Study:

  • To develop and evaluate a novel, rapid, and cost-effective point-of-care assay for Chlamydia trachomatis detection.
  • To assess the assay's feasibility using crude DNA lysis and loop-mediated isothermal amplification (LAMP) with hydroxynaphthol blue.

Main Methods:

  • Developed a one-step assay involving crude DNA lysis via heat treatment coupled with LAMP.
  • Utilized six primers targeting the C. trachomatis ompA gene for specificity.
  • Evaluated the assay on 284 endocervical swab specimens, comparing results to commercial DNA isolation and PCR methods.

Main Results:

  • The assay demonstrated high sensitivity (90-100%) and specificity (95%), comparable to conventional PCR.
  • Achieved detection of 0.04 pg of ompA DNA within a rapid 45-minute total assay time.
  • Results are visually interpreted via a color change (violet to sky blue) and the assay costs approximately $3 USD per reaction.

Conclusions:

  • The developed crude DNA lysis coupled LAMP assay is a simple, rapid, specific, sensitive, and cost-effective method for C. trachomatis detection.
  • This assay presents a viable alternative for clinical diagnosis in resource-limited healthcare facilities and field settings.
  • The assay utilizes safe reagents, ensuring user safety during testing.