Bacteria: Structure and Pathogenesis · USMLE Step 1

Why do Gram-positive bacteria retain the crystal violet stain but Gram-negative bacteria do not during Gram staining?

  1. Gram-negative bacteria completely lack peptidoglycan
  2. Alcohol dissolves the gram-negative outer membrane
  3. Gram-positive bacteria contain abundant LPS
  4. Gram-negative bacteria are rich in teichoic acids
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Correct answer: Alcohol dissolves the gram-negative outer membrane

During decolorization, alcohol/acetone disrupts the outer membrane of Gram-negative bacteria, making the thin peptidoglycan layer unable to retain the crystal violet–iodine complex. Gram-positive bacteria have thick peptidoglycan that traps the dye.

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