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Development of reverse transcription loop-mediated isothermal amplification-based assay for rapid and specific detection of human fungal pathogen, Candida auris

Summary

Researchers developed a new rapid test called RT-LAMP to detect Candida auris, a dangerous multidrug-resistant fungus that causes hospital infections. The test works by changing color from pink to yellow when the fungus is present, making it easy to read without special equipment. It is more sensitive and specific than current methods, can be done quickly at a single temperature, and costs much less than existing tests, making it ideal for hospitals and clinics with limited resources.

Background

Candida auris is a multidrug-resistant human fungal pathogen listed as critical by the WHO. It is frequently misidentified as other Candida species, leading to delayed diagnosis and treatment failures. Current diagnostic methods such as culture-based approaches are slow and less sensitive, while PCR-based methods are costly and unavailable in resource-limited settings.

Objective

This study aimed to develop and evaluate a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for rapid and specific detection of C. auris and compare its diagnostic efficiency with conventional LAMP assay using DNA as template.

Results

RT-LAMP achieved a limit of detection (LOD) of 1 attogram (ag) of RNA compared to 10 femtograms (fg) for LAMP using DNA, demonstrating superior sensitivity. The assay showed 100% specificity with no cross-reactivity to gram-negative bacteria or related Candida species. Clinical validation on 10 C. auris isolates demonstrated 100% concordance with culture-based methods.

Conclusion

RT-LAMP offers rapid, sensitive, and specific detection of C. auris with results visible by naked eye color change. The method is cost-effective, requires no specialized equipment, and shows promise for clinical application in resource-limited regions, though further large-scale clinical validation is warranted.
  • Published in:Current Medical Mycology,
  • Study Type:Methodology Development Study,
  • Source: PMID: 40584605
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