‫بسم هللا الرحمن الرحيم‬

‫‪Biochemical test‬‬
Biochemical test include :
• Catalase Test
• Oxidase Test
• Indole Ring Test
• MR Test
• VP Test
• Citrate Test
• Urease Test
• TSI Test
• Sugar Fermentation Test
• H2S Production Test
• Gelatin Hydrolysis Test
•
Catalase Test: Oxidase Test:
• is used to differentiate staphylococci (catalase-positive) • The oxidase test is used to determine if an organism
from streptococci (catalase-negative). possesses the cytochrome c oxidase enzyme.
Principle : Principle :
• 2H2O2 2H2O • Oxidase reagent ( colorless ) Indophenol ( purple color )
Example : Example :
No bubbles ( Negative ) Eg : Streptococcus sp Negative Eg : E. coli
Bubbles ( Positive )Eg : staphylococcus sp Positive Eg : Pseudomonas sp
Indole Ring Test: MR Test:
• The indole test screens for the ability of an organism to • MR Test: used for This test is used to determine which
degrade the amino acid tryptophan and produce indole. fermentation pathway is used to utilize glucose

Principle : Principle :
• tryptophan Utilize Indole Red complex • glucose Ferment
37 c
Pyruvic Acid Red in acidic pH
37 c
24-48 h 24-48 h

Example : Example :
Negative Eg : Salmonella sp Negative Eg : Klebsiella
Positive Eg : E. coli Positive Eg : E. coli
 VP Test: Citrate Test:
• VP test is used to determine if an organism produces • Citrate agar is used to test an organism's ability to
acetylmethyl carbinol from glucose fermentation. utilize citrate as a source of energy.
Principle : Principle :
• Glucose Acid acetylmethyl carbinol Red complex • Sod. Citrate (Neutral Ph 8.6) Carbonate,
Bicarbonate
Example : Example : Alkaline pH 7.6
Negative Eg : E. coli Negative Eg : E. coli
Positive Eg : Klebsiella Positive ( Blue) Eg : Salmonella sp

Note: If the MR test is positive, the VP test is negative and vice versa
Urease Test : Sugar Fermentation Test:
• It is primarily used to distinguish urease-positive • is used to determine whether or not a bacteria can
Proteeae from other Enterobacteriaceae. utilize a certain carbohydrate.

Principle : Principle :
• Urea NH3 + CO2 • Certain Sugar Acid Pink
Gas Bubble
Example : Example :
Negative Eg : E. coli Negative Eg : Salmonella sp
Positive Eg : staphylococcus Positive Eg : E. coli
H2S Production Test: TSI Test :
• Hydrogen sulfide is produced if the sulfur compound is • It is often used to differentiate enteric bacteria
reduced by the bacterial strain. including Salmonella

Principle :
• Ferrous Sulphate & Ferrous sulphide
Ferric Ammonium salt ( black Precipitate )

Example :
Negative Eg : shigella
Positive Eg : Salmonella sp
Gelatin Hydrolysis Test :
• The gelatin hydrolysis test detects the ability of bacteria to produce gelatinases.

Principle :
• Gelatin Amino acids ( Liquefy the Gelatin ) Liquefaction of Gelatin

Example :
Negative Eg : E. coli
Positive Eg : staphylococcus aureus