What test is used to diagnose onychomycosis (nail fungus)?

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From the Guidelines

To diagnose nail fungus, a fungal culture test or direct microscopic examination with Calcofluor white is typically used, as it provides the most accurate results. The diagnosis of nail fungus, also known as onychomycosis, requires laboratory testing to confirm the presence of a fungal infection 1. The clinical characteristics of dystrophic nails must alert the clinician to the possibility of onychomycosis, and laboratory detection and identification of dermatophytes consists of culture and microscopy 1.

Some key points to consider when diagnosing nail fungus include:

  • The use of Calcofluor white for direct microscopic examination of nail specimens, as it is exceedingly useful for seeing fungal elements 1
  • The importance of taking a small sample of the affected nail or debris beneath it and sending it to a laboratory to see if fungus grows 1
  • The need for an experienced microscopist and a laboratory with expertise in mycology to accurately interpret the results 1
  • The potential for incorrect diagnosis if laboratory testing is not used, as clinical diagnosis alone can be unreliable 1

In terms of specific tests, the following may be used:

  • Fungal culture test, which involves taking a small sample of the affected nail or debris beneath it and sending it to a laboratory to see if fungus grows 1
  • Potassium hydroxide (KOH) preparation, where nail scrapings are placed in a solution and examined under a microscope to identify fungal elements 1
  • Direct microscopic examination with Calcofluor white, which provides the most accurate results 1

From the FDA Drug Label

Prior to initiating treatment, appropriate nail specimens for laboratory testing (KOH preparation, fungal culture, or nail biopsy) should be obtained to confirm the diagnosis of onychomycosis. The tests used to see if there is nail fungus are:

  • KOH preparation
  • Fungal culture
  • Nail biopsy 2 2

From the Research

Diagnostic Tests for Nail Fungus

The following tests are used to diagnose nail fungus:

  • Direct microscopic examination with a potassium hydroxide (KOH) wet-mount preparation 3, 4, 5
  • Histopathologic examination of the trimmed affected nail plate with a periodic-acid-Schiff (PAS) stain 6, 3, 4, 5
  • Fungal culture 6, 7, 3, 4, 5
  • Polymerase chain reaction (PCR) assays 5

Sensitivity and Specificity of Diagnostic Tests

The sensitivity and specificity of these tests vary:

  • PAS staining is highly sensitive (93.1%) 3
  • KOH examination is highly sensitive but poorly specific 4
  • Fungal culture is highly specific but poorly sensitive 3, 4
  • Gomori's methenamine silver (GMS) stain is both highly sensitive and specific 4

Diagnostic Algorithm

A diagnostic algorithm for onychomycosis has been proposed, taking into consideration the sensitivity, cost-effectiveness, and necessary time for each test 3. The algorithm suggests using PAS staining as a primary diagnostic test, followed by fungal culture or KOH examination as secondary tests.

References

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Research

Onychomycosis: An Updated Review.

Recent patents on inflammation & allergy drug discovery, 2020

Research

Onychomycosis: Current trends in diagnosis and treatment.

American family physician, 2013

Research

Fungal nail infections: diagnosis and management.

Prescrire international, 2009

Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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