IMViC Tests: Principle, Procedure, Results

Written by Acharya Tankeshwar in Biochemical Tests Last Updated May 3, 2022

Each of the letters in “IMViC” stands for one of these tests. “I” is for indole; “M” is for methyl red; “V” is for Voges-Proskauer, and “C” is for citrate,  lowercase “i” is added for the ease of pronunciation. IMViC is an acronym that stands for four different tests

To obtain the results of these four tests, three test tubes are inoculated: tryptone broth (indole test), methyl red – Voges Proskauer broth (MR-VP broth), and citrate. IMViC tests are employed in the identification/differentiation of members of the family Enterobacteriaceae.

General procedure for performing IMViC Tests and their interpretations:

Cultures of any members of Enterobacteriaceae have to grow for 24 to 48 hours at 37°C and the respective tests can be performed:

Indole test

It is performed on sulfide-indole-motility (SIM) medium or in tryptophan broth, or in motility urease indole (MIU) medium. Result is read after adding Kovac’s reagent.

  1. The positive result is indicated by the red layer at the top of the tube after the addition of Kovács reagent.
  2. A negative result is indicated by the lack of color change at the top of the tube after the addition of Kovács reagent.
indole test results
Indole Test Results: Positive-development of Red-ring

Methyl red test and Voges-Proskauer test both are done in methyl red–Voges-Proskauer (MR-VP) broth, but the reagents that are added varies according to the tests.

Methyl Red (MR) test

  • Positive methyl red test is indicated by the development of red color after the addition of methyl red reagent.
  • A negative methyl red test is indicated by no color change after the addition of methyl red reagent

Voges-Proskauer (VP) test

  1. Negative VP test is indicated by a lack of color change after the addition of Barritt’s A and Barritt’s B reagents.
  2. A positive Voges-Proskauer test is indicated by the development of red-brown color after the addition of Barritt’s A and Barritt’s B reagents.

 Citrate Utilization test

The test is performed on Simmons citrate agar:

  1. Negative citrate utilization test is indicated by the lack of growth and color change in the tube
  2.  A positive citrate result is indicated by growth and a blue color change. 

IMViC Test results of some genera of Enterobacteriaceae:       

IMViC tests of Escherichia coli

  • Indole: Positive
  • Methyl-Red: Positive
  • Voges-Proskauer test: Negative
  • Citrate test: Negative
IMViC Test of Ecoli: ++--
IMViC Test of Ecoli: ++– (Photo source and  copyright: ASM)

IMViC tests of Klebsiella (formerly Enterobacter) aerogenes

  1. Indole: Negative
  2. Methyl-Red: Negative
  3. Voges-Proskauer test: Positive
  4. Citrate test: Positive

IMViC tests of Proteus vulgaris

  1. Indole: Positive
  2. Methyl-Red: Positive
  3. Voges-Proskauer test: Negative
  4. Citrate test: Negative

IMViC tests of Citrobacter freundii

  1. Indole: Negative
  2. Methyl-Red: Positive
  3. Voges-Proskauer test: Negative
  4. Citrate test: Positive

Acharya Tankeshwar

Hello, thank you for visiting my blog. I am Tankeshwar Acharya. Blogging is my passion. As an asst. professor, I am teaching microbiology and immunology to medical and nursing students at PAHS, Nepal. I have been working as a microbiologist at Patan hospital for more than 10 years.

23 thoughts on “IMViC Tests: Principle, Procedure, Results”

  15. Something to note about the tests mentioned:
    SIM test, its is 3 tests in one:
    Indole
    Motility
    Hydrogen sulfide production

    http://www.microbelibrary.org/library/2-associated-figure-resource/3488-sulfur-indole-motility-test-results-from-various-microbes

    MR/VP testing is done with one media for 2 test. Grow the organism in the broth then remove 1 ml to a clean test tube to do the vp test

    http://www.microbeworld.org/component/jlibrary/?view=article&id=12634

    Citrate, organism uses nitrate as sole source of carbon:

    http://www.microbelibrary.org/library/2-associated-figure-resource/2534-citrate-test-on-escherichia-coli-and-enterobacter-aerogenes

    Reply

    1. Hello Tasha
      I was reading your comments and found we failed to reply some of them and acknowledge the others, sorry for that. In this particular comment two links are not working (may be url has been changed after microbelibary website is revamped).

      Reply

      1. Try these links:
        http://www.microbeworld.org/component/jlibrary/?view=article&id=12601
        and
        http://www.microbeworld.org/component/jlibrary/?view=article&id=12634
        Cheers,
        Tasha

        Reply

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