Kovac's Indole Reagent: Intended Use

Kovac's Indole Reagent R008
Intended use
For detection of presence of indole produced by microorganisms due to tryptophan deamination.
Composition**
Ingredients -
p-dimethylamino benzaldehyde 5.000
Amyl alcohol 75.000
Hydrochloric acid, concentrated 25.000
**Formula adjusted, standardized to suit performance parameters
Directions
Add 0.2 - 0.3 ml of Kovac's reagent to 5 ml of a 24 - 48 hours old culture of the organism under investigation. Formation of
a red coloured ring indicates positive indole test.
Principle And Interpretation
Peptone Water is particularly suitable as a substrate in the study of indole production. Peptone used in Peptone Water, is rich
in tryptophan content (1). Other peptones which contain tryptophan can be used to study indole production. Tryptone Water
is also recommended by APHA (2) for detection of indole production by coliforms, which is a key feature in differentiation
of bacteria. It is used as part of the IMViC procedures. Most strains of E. coli, P. vulgaris, P. rettgeri, M. morgani and
Providencia species break down the amino acid tryptophan with the release of indole. The presence of indole can be detected
by the addition of Ehrlich's or Kovac's reagent (p-dimethylaminobenzaldehyde).
Kovacs reagent is a biochemical reagent consisting of isoamyl alcohol, para-dimethylaminobenzaldehyde (DMAB), and
concentrated hydrochloric acid. It is used for the diagnostic test, to determine the ability of the organism to split tryptophan
into indole and alpha-aminopropionic acid by hydrolytic activity of bacteria that express tryptophanase enzyme (3). The
indole produced is indicated by formation of a red coloured ring, soluble in ether, chloroform and alcohol. This was invented
by the Hungarian-Swiss Chemist, Ervin Kovats. Indole production is used as, a tests designed to distinguish among members
of the family Enterobacteria.
Type of specimen
Clinical samples ; Water samples
Specimen Collection and Handling:
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (4,5).
For water samples, follow appropriate techniques for sample collection, processing as per guidelines and local standards(2).
After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions :
In Vitro diagnostic Use only. Read the label before opening the container. Wear protective gloves/protective clothing/eye
protection/ face protection. Follow good microbiological lab practices while handling specimens and culture. Standard
precautions as per established guidleines should be followed while handling clincal specimens. Saftey guidelines may be
referred in individual safety data sheets
Limitations :
1. Growth media must contain an adequate amount of tryptophan. Do not use Mueller- Hinton Agar for test, because
tryptophan is destroyed during the acid hydrolysis of casein.
2. Do not used media that contain dyes (e.g., EMB, MAC).
3. Do not use medium with a nitrate disc/strip to perform the indole test, as nitrate can interfere with indole test by
including false positive results.
Performance and Evaluation
Performace of the product is expected when used as per the direction on the label within the expiry period when stored at
recommended temperature.
Please refer disclaimer Overleaf.

HiMedia Laboratories Technical Data
Quality Control
Appearance
Greenish yellow coloured solution
Solubility
Immiscible with water
Clarity
Clear with no insoluble particles.
Cultural Response
Characteristic reactions observed when Kovac's Indole Reagent is added to growth in Tryptone Broth (M463).
Organism Indole production
Enterobacter aerogenes ATCC 13048 negative reaction ,no red ring
Escherichia coli ATCC 25922 positive reaction, red ringreaction, red ring at the interface of the medium
Storage and Shelf Life
Store between 10-30°C in tightly closed container and away from bright light. Use before expiry date on label. On opening,
product should be properly stored in dry ventilated area protected from extremes of temperature and sources of ignition.
Seal the container tightly after use.
Disposal
User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. Follow
established laboratory procedures in disposing of infectious materials and material that comes into contact with clinical
sample must be decontaminated and disposed of in accordance with current laboratory techniques (4,5).
Reference
1.MacFaddin J., 1980, Biochemical Tests for Identification of Medical Bacteria, 2nd ed., Williams and Wilkins, Baltimore.
2.Greenberg A. E., Clesceri L. S. and Eaton A. D., (Eds.), 2005, Standard Methods for the Examination of Water and
Wastewater, 21st ed., APHA, Washington, D.C.
3.MacFaddin J. F., 2000, Biochemical Tests for Identification of Medical Bacteria, 3rd Ed., Williams and Wilkins, Baltimore.
4.Isenberg, H.D. Clinical Microbiology Procedures Handb0ook. 2nd Edition.
5. Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015)
Manual of Clinical Microbiology, 11th Edition. Vol. 1.
Revision :01 / 2017

In vitro diagnostic
IVD
medical device
CE
Marking
30° Storage
C
10° temperature
C
Do not use if
package is
damaged
HiMedia Laboratories Pvt.
Limited, B /4-6 , MIDC,
Dindori, Nashik MH
www.himedialabs.com
EC REP CE Partner 4U ,Esdoornlaan

13, 3951 DB Maarn The
Netherlands,
www.cepartner 4u.eu
Disclaimer :
User must ensure suitability of the product(s) in their application prior to use. Products conform solely to the information contained in
this and other related HiMedia™ publications. The information contained in this publication is based on our research and development
work and is to the best of our knowledge true and accurate. HiMedia™ Laboratories Pvt Ltd reserves the right to make changes to
specifications and information related to the products at any time. Products are not intended for human or animal or therapeutic use but
for laboratory,diagnostic, research or further manufacturing use only, unless otherwise specified. Statements contained herein should not
be considered as a warranty of any kind, expressed or implied, and no liability is accepted for infringement of any patents.
HiMedia Laboratories Pvt. Ltd. Reg.office : 23, Vadhani Ind.Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6116 9797 Corporate office :
A-516,Swastik Disha Business Park,Via Vadhani Ind. Est., LBS Marg, Mumbai-400086, India. Customer care No.: 022-6147 1919 Email:
techhelp@himedialabs.com Website: www.himedialabs.com

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Revision :01 / 2017

EC REP CE Partner 4U ,Esdoornlaan

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