Microbiology · Healthcare-Associated Infections and Hospital Microbiology (CLABSI, CAUTI, VAP, Sterilization Monitoring)

For monitoring the efficacy of an autoclave (steam sterilization), which biological indicator is used, and what is its specific endpoint test?

  • A Bacillus atrophaeus spores — used for dry heat sterilization monitoring, not autoclaving
  • B Geobacillus stearothermophilus spores — positive (yellow/turbid) broth after incubation indicates sterilization failure
  • C Clostridium sporogenes spores — anaerobic indicator that changes colour in aerobic conditions
  • D Aspergillus niger spores — fungal indicator most resistant to steam
Correct answer: B. Geobacillus stearothermophilus spores — positive (yellow/turbid) broth after incubation indicates sterilization failure

Explanation

The biological indicator for autoclave (moist heat/steam sterilization) is Geobacillus stearothermophilus (formerly Bacillus stearothermophilus) spores; the standard run is 121°C for 15 minutes (gravity cycle) or 132°C for 4 minutes (pre-vacuum). After autoclaving, the indicator strip is incubated at 56°C for 24–48 hours; if the pH indicator in the broth remains purple (no acid production, no growth), sterilization was successful. Yellow/turbid broth indicates surviving spores and sterilization failure. Bacillus atrophaeus is used for ethylene oxide and dry heat monitoring.

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

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