KB003
KB003
KB003
Principle
Each Hi25™ kit is a standardized colorimetric identification system utilizing thirteen conventional biochemical tests and eleven carbohydrate utilization tests. The tests are based on
the principle of pH change and substrate utilization. On incubation organisms undergo metabolic changes which are indicated by a colour change in the media that is either visible
spontaneously or after addition of a reagent. Oxidase test is performed separately using oxidase reagent disc provided with the kit.
Kit contents
1. Each kit contains sufficient material to perform 10 tests. 8. TDA reagent (R036) for Phenylalanine Deaminase test.
2. 10 kits of Part I. 9. Baritt reagent A (R029) for Voges-Proskauer's test.
3. 10 kits of Part II. 10. Baritt reagent B (R030) for Voges-Proskauer's test.
4. Oxidase reagent discs (DD018) 11. Methyl Red reagent (I007) for Methyl Red test
5. Technical product insert. 12. Kovac's reagent (R008) for Indole test
6. Result Interpretation Chart and Result Entry Datasheet. 13. Sulphanilic acid (R015)
7. Identification Index. 14. N,N-Dimethyl-1-Napthylamine Reagent (R009).
Interpretation of results :
Interpret results as per the standards given in the identification index. Addition of reagents wherever required should be done at the end of incubation period that is after
18 - 24 hours.
Part I : Phenylalanine Deamination Test : Well No. 5
Add 2-3 drops of TDA reagent (R036). Development of dark green colour within one minute indicates a positive reaction.
No change in colour denotes a negative reaction.
Nitrate Reduction Test : Well No. 6 Add 1-2 drops of Sulphanilic acid (R015) and 1-2 drops of N,N-Dimethyl-1-Napthylamine Reagent (R009).
Immediate development of pinkish red colour on addition of reagent indicates positive reaction. No change in colour indicates a negative reaction.
Voges Proskauer's Test : Well No. 9 Add 2-3 drops of Baritt reagent A (R029) and 1 drop of Baritt reagent B(R030).
Pinkish red colour development within 5-10 minutes indicates a positive test.
No change in colour or a slight change in colour (due to reaction of Baritt reagent A with Baritt reagent B) denotes a negative reaction.
Methyl Red Test : Well No. 10 Add 1-2 drops of Methyl Red reagent (I007).
Reagent remains red in colour if the test is positive.
Reagent decolourises and becomes yellow if the test is negative.
Indole Test : Well No. 11 Add 1-2 drops of Kovac's reagent (R008).
Development of pinkish red colour within 10 seconds indicates positive reaction.
Reagent remains pale coloured if the test is negative.
Tests ONPG Lysine Ornithine Urease TDA Nitrate H2S Citrate Utilization Voges Proskauer's Methyl Red Indole Malonate
Budvicia aquatica
Buttiauxella agrestis
Cedecea davisae
Cedecea lapagei
Cedecea neteri
Citrobacter amalonaticus
Citrobacter diversus
Citrobacter freundii
Enterobacter aerogenes
Enterobacter amnigenus (Biogroup I)
Enterobacter amnigenus (Biogroup II)
Enterobacter taylorae (E. cancerogenus)
Enterobacter cloacae
Enterobacter gergoviae
Enterobacter sakazakii
Escherichia coli
Escherichia coli, inactive
Escherichia blattae
Escherichia fergusonii
Escherichia hermannii
Escherichia vulneris
Ewingella americana
Hafnia alvei
Klebsiella oxytoca
Klebsiella pneumoniae
subspecies ozaenae
Proteus vulgaris
Providencia alcalifaciens
Providencia rettgeri
Providencia rustigianii
Rahnella aquatilis
Salmonella bongori
Salmonella choleraesuis subspecies arizonae
Salmonella choleraesuis
subspecies choleraesuis
Salmonella choleraesuis subspecies diarizonae
Salmonella choleraesuis subspecies houtenae
Salmonella choleraesuis subspecies indica
Salmonella choleraesuis subspecies salamae
Salmonella enteritidis
Salmonella typhi
Salmonella typhimurium
Serratia entomophila
Serratia ficaria
Serratia fonticola
Serratia marcescens
Serratia odorifera (Biogroup I)
Serratia odorifera (Biogroup II)
Serratia plymuthica
Serratia proteamaculans
Serratia rubidaea
Shigella boydii, Shigella flexneri,
Shigella dysenteriae
Shigella sonnei
Yersinia enterocolitica
Yersinia frederiksenii
Yersinia intermedia
Yersinia pestis
Yersinia pseudotuberculosis
Note : Based on % strains showing reactions following symbols have been assigned from laboratory results and standard references.
+ = Positive (more than 90%) = Negative (more than 90%) V = 11-89% positive.
Esculin hydrolysis Arabinose Xylose Adonitol Rhamnose Cellobiose Melibiose Saccharose Raffinose Trehalose Glucose Lactose
Note : Based on % strains showing reactions following symbols have been assigned from laboratory results and standard references.
+ = Positive (more than 90%) = Negative (more than 90%) V = 11-89% positive.
5 Phenylalanine 2-3 drops of TDA reagent Detects Phenylalanine Colourless Green Colourless
Deamination deamination activity
6 Nitrate 1-2 drops of sulphanilic Detects Nitrate reduction Colourless Pinkish Red Colourless
reduction acid and 1-2 drops of N,
N-Dimethyl-1-Napthylamine
7 H2S production — Detects H2S production Orangish yellow Black Orangish yellow
9 Voges 1-2 drops of Baritt reagent A Detects acetoin production Colourless / Light Pinkish red Colourless/
Proskauer's and 1-2 drops of Baritt reagent B Yellow slight copper
10 Methyl red 1-2 drops of Methyl red reagent Detects acid production Colourless Red Yellowish- orange
11 Indole 1-2 drops of Kovac's red reagent Detects deamination of tryptophan Colourless Pinkish Red Colourless
1. Allow the reagents to come to room temperature after removal from the refrigerator .
2. In case of carbohydrate fermentation test some microorganisms show weak reaction. In this case record the reaction as ± and incubate further for 48 hours.
Orange colour after 48 hours of incubation should be interpreted as a negative reaction.
3. In case of Lysine and Ornithine decarboxylation, incubation up to 48 hours may be required.
4. At times organisms give conflicting result because of mutation or the media used for isolation, cultivation and maintenance.
5. The identification index has been compiled from standard references and results of tests carried out in the laboratory.
Precautions :
Clinical samples and microbial cultures should be considered potentially pathogenic and handled accordingly.
Aseptic conditions should be maintained during inoculation and handling of the kits.
Reagents should not come in contact with skin, eyes or clothing.
After use, kits and the instruments used for isolation and inoculation (pipettes, loops etc.) must be disinfected using a suitable disinfectant and then discarded by
incineration or autoclaving in a disposal bag.
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 diagnostic or therapeutic use but for
laboratory, 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. 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