Microbiology Lecture for MBBS Students
Urease Test
Principle
• • Detects ability of microorganisms to produce
urease enzyme.
• • Urease hydrolyzes urea → ammonia (NH₃) +
carbon dioxide (CO₂).
• • Ammonia increases pH → medium becomes
alkaline.
• • Phenol red indicator:
• - Yellow = acidic (pH < 6.8)
• - Red = neutral
• - Pink (fuchsia) = alkaline (pH > 8.1, urease
positive).
Medium
• • Christensen’s Urea Agar slant (commonly
used)
• - Contains urea, peptones, glucose, phenol
red
• • Stuart’s Urea Broth (more sensitive, for
fastidious organisms)
Procedure
• 1. Inoculate the test organism onto Urea agar
slant or into Urea broth.
• 2. Incubate at 35–37°C for 24–48 hrs (some
organisms up to 7 days).
• 3. Observe color change.
Results & Interpretation
• • Positive test: Pink (fuchsia) color within 24–48
hrs → urease produced.
• • Negative test: No color change (medium
remains yellow/orange) → no urease.
Examples
•• Positive organisms:
• - Proteus spp. (rapid positive)
• - Klebsiella pneumoniae
• - Helicobacter pylori
• - Cryptococcus neoformans (yeast)
•• Negative organisms:
• - Escherichia coli
• - Salmonella spp.
• - Shigella spp.
Precautions
• • Use light inoculum (avoid false positives).
• • Incubation time is critical:
• - Rapid urease producers: few hours (Proteus)
• - Weak producers: 24–48 hrs (Klebsiella)
• • Avoid over-incubation (false results).
• • Use fresh medium (urea may degrade
spontaneously).