Precision diagnosis: Onychomycosis detection with multiplex real-time PCR versus conventional methods

Ramadan, Amira E.; Mohamed, Miran K.; El-sayed, Marwa S.; Nassar, Nora M.; Elsayed, Rasha A.

doi:10.21608/mid.2025.362150.2562

	Precision diagnosis: Onychomycosis detection with multiplex real-time PCR versus conventional methods
Microbes and Infectious Diseases
Articles in Press, Accepted Manuscript, Available Online from 28 March 2025
Document Type: Original Article
DOI: 10.21608/mid.2025.362150.2562
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Authors
Amira E. Ramadan ¹; Miran K. Mohamed²; Marwa S. El-sayed³; Nora M. Nassar⁴; Rasha A. Elsayed¹
¹Department of Medical Microbiology and Immunology, Faculty of Medicine, Benha University, Egypt
²Department of Dermatology, venereology and Andrology, Faculty of Medicine, Benha University, Egypt,
³Department of Medical Microbiology and Immunology, Faculty of Medicine, Benha University, Egypt,
⁴Department of clinical and chemical pathology, Faculty of Medicine, Benha University, Egypt
Abstract
Background: One of the most prevalent nail conditions is onychomycosis, that is mostly brought on by dermatophyte fungi, non-dermatophyte molds (NDMs) and yeast which may be confused with other nail lesions so, it is crucial to accurately identify the causative agent to initiate proper medications. Fungal culture and KOH microscopy have always been the gold standards for diagnosis. Aim: This study aimed to compare the results of a multiplex real-time PCR (RT-PCR) assay with those of conventional diagnostic methods (direct microscopy and culture) in detecting and diagnosing onychomycosis. Methods: A total of 100 nail samples from clinically suspected onychomycosis patients were divided into 3 pieces, one nail piece was set aside for microscopy, another for culture and the rest were kept at room temperature in sterile screwed vials in preparation for the multiplex RT-PCR test and DNA extraction. Results: of 100 nail samples, 30 had negative direct KOH and 70 had positive direct KOH. In the group of negative KOH samples, 10/30 had a positive culture. In the group of positive K OH samples, 70/100 had a positive culture. Multiplex RT-PCR demonstrated 92.8% sensitivity, 62.7% specificity, 61.9% VPP and 93.1% VPN. Conclusion: using multiplex RT-PCR can improve the detection of onychomycosis and decrease the turn round time which could improve the disease outcome.
Keywords
RT-PCR; KOH; NDMs


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