When “Clean” Isn’t Clean: A Diagnostic Pitfall in Gram Staining Leading to Pseudo-Bacterial Meningitis

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Abstract

Introduction: The Gram stain is a fundamental and widely used technique in clinical microbiology for its simplicity and diagnostic value, particularly in bacterial meningitis. However, its reliability can be compromised by contamination and artifacts. Case Presentation: This report describes the case of pseudo-bacterial meningitis in a previously healthy 25-year-old woman who presented with fever, headache, and nuchal rigidity. Cerebrospinal fluid (CSF) analysis revealed pleocytosis with mononuclear predominance. Gram staining demonstrated structures consistent with gram-negative bacilli, prompting empiric antibacterial therapy, with concurrent antiviral treatment. Comprehensive microbiological investigations, including CSF culture and broad-range 16S rRNA PCR, yielded negative results. Residual materials or microscopic debris on supposedly “clean” slides produced artifact-like appearances that mimicked microorganisms, leading to false-positive interpretations. Conclusions: This case highlights the critical importance of rigorous laboratory practices, including proper slide preparation and procedural standardization, as well as awareness of potential artifacts to prevent misdiagnosis.

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