What laboratory tests are essential for confirming a diagnosis of diphtheria in a suspected case?

Guideline-aligned answer with reasoning, red flags and references. Clinically reviewed by Dr Kola Tytler MBBS CertHE MBA MRCGP.

Posted: 22 August 2025Updated: 22 August 2025 Guideline-Aligned (High Confidence) Clinically Reviewed
Dr Kola Tytler MBBS CertHE MBA MRCGPClinical Lead • iatroX

Essential laboratory tests for confirming a diagnosis of diphtheria include:

  • Culture of throat or lesion swabs on selective media to isolate Corynebacterium diphtheriae or C. ulcerans, which is the primary diagnostic step .
  • Microscopic examination of the specimen using Albert’s stain or Gram stain to identify characteristic gram-positive bacilli with metachromatic granules supports the diagnosis .
  • Biochemical identification and toxigenicity testing of the isolated strain are crucial to confirm the presence of toxigenic C. diphtheriae, as only toxigenic strains cause diphtheria .
  • Detection of diphtheria toxin production is essential and can be performed by the Elek immunodiffusion test, which remains the gold standard for confirming toxigenicity .
  • Molecular methods such as PCR targeting the tox gene can rapidly detect toxigenic strains and complement culture methods, especially when toxin expression is low or culture is negative .
  • Recent literature highlights challenges in toxin detection and suggests that combining culture, tox gene PCR, and Elek test improves diagnostic accuracy .

In summary, the essential laboratory confirmation of diphtheria requires isolation of the organism from clinical specimens, demonstration of toxigenicity by Elek test or molecular methods, and biochemical identification to differentiate toxigenic strains .

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