IMViC


The IMViC tests are a group of individual tests used in microbiology lab testing to identify an organism in the coliform group. A coliform is a gram negative, aerobic, or facultative anaerobic rod, which produces gas from lactose within 48 hours. The presence of some coliforms indicate fecal contamination.
The term "IMViC" is an acronym for each of these tests. "I" is for indole test; "M" is for methyl red test; "V" is for Voges-Proskauer test, and "C" is for citrate test. The lower case "i" is merely for "in" as the Citrate test requires coliform samples to be placed "in Citrate". Triple sugar iron test, Urease Test and Malonate Test are also included in this test.
These tests are useful in distinguishing members of Enterobacteriaceae.

Indole test

In this test, the organism under consideration is grown in peptone water broth. It contains tryptophan, which under the action of enzyme tryptophanase is converted to an Indole molecule, pyruvate and ammonium. The indole is then extracted from the broth by means of xylene. To test the broth for indole production, Kovac's reagent. Kovac's reagent consist of amyl alcohol and para-dimethylaminobenzaldehyde and concentrated hydrochloric acid. Kovac's reagent is actually used to determine ability of an organism to separate indole from amino acid tryptophan and it is added after incubation. A positive result is indicated by a pink/red layer forming on top of the liquid.

Methyl red and Voges–Proskauer test

These tests both use the same broth for bacterial growth. The broth is called MRVP broth. After growth, the broth is separated into two different tubes, one for the methyl red test and one for the Voges-Proskauer test.
The methyl red test detects production of acids formed during metabolism using mixed acid fermentation pathway using pyruvate as a substrate. The pH indicator Methyl Red is added to one tube and a red color appears at pH's lower than 4.2, indicating a positive test. The solution remaining yellow indicates a negative test, meaning the butanediol fermentation is used.
The VP test uses alpha-naphthol and potassium hydroxide to test for the presence of acetylmethylcarbinol, an intermediate of the 2,3-butanediol fermentation pathway. After adding both reagents, the tube is shaken vigorously then allowed to sit for 5-10 minutes. A pinkish-red color indicates a positive test, meaning the 2,3-butanediol fermentation pathway is used.

Citrate test

This test uses Simmon's citrate agar to determine the ability of a microorganism to use citrate as its sole carbon source. The agar contains citrate and ammonium ions and bromothymol blue as an indicator. The citrate agar is green before inoculation, and turns blue as a positive test indicator, meaning citrate is utilized.

Usage

These IMViC tests are useful for differentiating the family Enterobacteriaceae, especially when used alongside the Urease test.
The IMViC results of some important species are shown below.
SpeciesIndoleMethyl RedVoges-ProskauerCitrate
Escherichia coliPositivePositiveNegativeNegative
Shigella spp.NegativePositiveNegativeNegative
Salmonella spp.NegativePositiveNegativePositive
Klebsiella spp.NegativeNegativePositivePositive
Proteus vulgarisPositivePositiveNegativeNegative
Proteus mirabilisNegativePositiveNegativePositive
Citrobacter freundiiNegativePositiveNegativePositive
Enterobacter aerogenesNegativeNegativePositivePositive