Indole test Principle and Protocol

Indole test Principle and Protocol:

Indole is a hetrocyclic aromatic compound, which is usually a product of degeneration of tryptophan. 

Enzyme which plays a crucial role on breaking down tryptophan is called 'Tryptophanase.'

Basic reaction for breakdown of Indole:
Water + Tryptophan --[Tryptophanase]--> Indole + Pyruvate + Ammonia

Pyruvate is used for respiration or fermentation processes. 



Principle: Bacteria break down tryptophan to indole which is detected by adding Kovac's reagent which gives a cherry red color on reaction with indole.

Kovac's reagent is used to diagnose this reaction. Kovac's reagent consists of isoamyl alcohol, para-dimethylaminobenzaldehyde and concentrated hydrochloric acid.

Indole + Kovac's reagent ---> Cherry Red Color

Indole positive organisms: Escherichia coli, Haemophilus influenzae, Streptococcus faecalis, Vibrio sp.

Indole negative organisms: Proteus mirabilis, Lactobacillus spp. , Pseudomonas sp., Salmonella sp.