Microbiology · Mycobacteria (Tuberculosis, Leprosy, Atypical)

A sputum smear from a suspected TB patient is reported as 3+ AFB positive on Ziehl-Neelsen staining. In Ziehl-Neelsen staining, the primary stain used and the reason for acid-fastness of mycobacteria is:

  • A Crystal violet; retained due to thick peptidoglycan layer
  • B Methylene blue; retained due to lipopolysaccharide in outer membrane
  • C Carbol fuchsin; retained due to high mycolic acid content in the cell wall
  • D Safranin; retained due to waxy coat preventing decolorization
Correct answer: C. Carbol fuchsin; retained due to high mycolic acid content in the cell wall

Explanation

Ziehl-Neelsen staining uses carbol fuchsin as the primary stain, with heat application (hot method) to drive the stain into the bacteria. Mycobacteria are acid-fast because their cell walls contain large amounts of mycolic acids (long-chain branched fatty acids) combined with arabinogalactan and peptidoglycan. This waxy lipid-rich wall resists decolorization by 3% hydrochloric acid in alcohol (acid-alcohol decolorizer), and the bacteria retain the red carbol fuchsin dye against the blue methylene blue counterstain. Fluorescent staining with auramine-rhodamine has higher sensitivity for screening.

Reference: Ananthanarayan & Paniker's Textbook of Microbiology, 11th ed.

High-yield for: NEET PGINI-CETNExTFMGEUSMLEPLABMRCP

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