Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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HiIMViC™ Biochemical Test Kit
KB001 is a standardized, colorimetric identification system utilizing twelve conventional biochemical tests for identification and differentiation of Enterobacteriaceae species using pure isolate. Organisms belonging to Enterobacteriaceae are Gram negative, oxidase negative, nitrate positive rods and are the most frequently isolated bacteria from clinical specimen and non clinical samples.
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Kit contents
- Each kit contains 5/10/20 kits of KB001, sufficient material to perform 5/10/20 tests.
• (a combination of 12 tests for differentiation of Enterobacteriaceae species. (Kit contains sterile media for Indole, Methyl red, Voges Proskauer's, Citrate utilization tests and 8 different carbohydrates- Glucose, Adonitol, Arabinose, Lactose, Sorbitol, Mannitol, Rhamnose, Sucrose) - Technical product insert.
- Identification Index.
- Kovac's Reagent (R008) for Indole Test,
- Methyl Red Indicator (1007) for MR test
- Baritt reagent A (R029) for VP test
- Baritt reagent B (R030) for VP test
- Result Interpretation Chart and Result Entry Datasheet.
Material Required but not supplied
- McFarland standard
- Inoculation loops, pipettes
- Enrichment medium / Isolation media
Direction
Preparation of inoculum :
- Isolate the organism to be identified on a common medium like Nutrient Agar (M001) or Tryptone Soya Agar (M290).
- Pick up a single isolated colony and inoculate in 5 ml BHI Broth (M210) and incubate at 35-37°C for 4-6 hours until the inoculum turbidity is 0.1 OD at 620nm or 0.5 McFarland standard. Some organisms may require more than 6 hours of incubation. In this case incubate till the inoculum turbidity reaches 0.1 OD at 620nm.
- Alternatively, prepare the inoculum by picking 1-3 well isolated colonies and make a homogenous suspension in 2-3 ml sterile saline. The density of the suspension should be 0.1 OD at 620nm.
Inoculation of the kit :
- Open the kit aseptically. Peel off the sealing foil.
- Inoculate each well with 50 µl of the above inoculum by surface inoculation method.
- Alternatively, the kit can also be inoculated by stabbing each individual well with a loopful of inoculum
Incubation:
Temperature of incubation: 35 - 37°C. Duration of incubation: 18 - 24 hours
Interpretation of results :
Interpret results as per the standards given in the identification index. Addition of reagents in well no 1,2 and 3 should be done at the end of incubation period that is after 18 - 24 hours.
Principle
KB001 is a standardized, colorimetric identification system utilizing twelve conventional biochemical tests. The tests are based on the principle of pH change and substrate utilization. On incubation, organisms undergo metabolic changes which are indicated as a colour change in the media that is either visible spontaneously or after addition of a reagent (1).
Type of specimen
Pure isolate from clinical specimen and non clinical sample
Specimen collection and handling
Refer direction
Warning and Precautions
In Vitro diagnostic Use. For professional use only. Read the label before opening the pack. Wear protective gloves/ protective clothing/eye protection/ face protection. Follow good microbiological lab practices while handling specimens and culture. Standard precautions as per established guidelines should be followed while handling clinical specimens. Aseptic conditions should be maintained during inoculation and handling of the kits. Reagents should not come in contact with skin, eyes or clothing. Safety guidelines may be referred in individual safety data sheets.
Limitations
- Allow the reagents to come to room temperature after removal from the refrigerator.
- In case of carbohydrate fermentation test some microorganisms show weak reaction. In this case record the reaction as ± and incubate further upto 48 hours. Orange colour after 48 hours of incubation should be interpreted as a negative reaction.
- At times organisms give conflicting result because of mutation or the media used for isolation, cultivation and maintenance.
- The identification index has been compiled from standard references and results of tests carried out in the laboratory.
- Erroneous false negative results may be obtained if the inoculum turbidity is less than 0.1 O D.
- Results are more prominent if an enriched culture is used instead of a suspension.
- It cannot be used directly for clinical specimens. The microorganisms to be identified have to be first isolated on appropriate isolation media. Only pure cultures should be used.
Performance and Evaluation
Performance of the medium is expected when used as per the direction on the label within the expiry period when stored at recommended temperature.
Quality Control
Appearance
Sterile white opaque strip with 12 wells containing media for 1.Indole, 2.Methyl red, 3. Voges Proskauer's, 4.Citrate utilization tests and 8 different carbohydrates- 5. Glucose, 6.Adonitol, 7. Arabinose, 8.Lactose, 9.Sorbitol, 10.Mannitol, 11.Rhamnose, 12.Sucrose
Quantity of medium
0.8 ml of medium in each well.
Sterility Check
Passes release criteria
Interpretation of results :
Interpret results as per the standards given in the identification index. Addition of reagents in no.1,2 and 3 well and should be done at the end of incubation period that is after 18 - 24 hours.
- Indole Test: Well No. 1
- Add 2-3 drops of Kovac's reagent (R008).
- Positive test is indicated by a development of pinkish red ring.
- No red ring indicates a negative reaction.
- Methyl red Test: Well No. 2
- Add 2-3 drops of Methyl red indicator (I007).
- Positive test is indicated by a development of red colour.
- Yellowish orange colour indicates a negative reaction.
- Voges-Proskaeur's Test : Well No. 3
- Add 2-3 drops of Baritt reagent A (R029) and 1 drop of Baritt reagent B (R030).
- Positive test is indicated by a development of pinkish red colour in 5 - 10 minutes.
- No colour change or a copper colour (due to reaction of Reagent A and Reagent B) indicates a negative reaction.
- Citrate Test: Well No. 4
- Positive test is indicated by a colour change to blue colour.
- Green or no colour change indicates a negative reaction.
- Carbohydrate utilization Test: Well No. 5-12
- Positive test is indicated by a colour change to yellow colour.
- Red or no colour change indicates a negative reaction.
Result Interpretation chart
| No. | Test | Reagents to be added after incubation | Principle | Original colour of the medium | Positive reaction | Negative reaction |
|---|---|---|---|---|---|---|
| 1 | Indole | 1-2 drops of Kovac's reagent | Detects deamination of tryptophan | Colourless | Reddish pink | Colourless |
| 2 | Methyl red | 1-2 drops of Methyl red indicator | Detects acid production | Colourless/ light yellow | Red | Yellowish - orange |
| 3 | Voges Proskauer's | 1-2 drops of Baritt reagent A and 1-2 drops of Baritt reagent B | Detects acetoin production | Colourless / light yellow | Pinkish red | Colourless/ slight copper |
| 4 | Citrate utilization | — | Detects capability of organism to utilize citrate as a sole carbon source | Green | Blue | Green |
| 5 | Glucose | — | Glucose utilization | Pinkish Red / Red | Yellow | Red |
| 6 | Adonitol | — | Adonitol utilization | Pinkish Red / Red | Yellow | Red |
| 7 | Arabinose | — | Arabinose utilization | Pinkish Red / Red | Yellow | Red |
| 8 | Lactose | — | Lactose utilization | Pinkish Red / Red | Yellow | Red |
| 9 | Sorbitol | — | Sorbitol utilization | Pinkish Red / Red | Yellow | Red |
| 10 | Mannitol | — | Mannitol utilization | Pinkish Red / Red | Yellow | Red |
| 11 | Rhamnose | — | Rhamnose utilization | Pinkish Red / Red | Yellow | Red |
| 12 | Sucrose | — | Sucrose utilization | Pinkish Red / Red | Yellow | Red |
Result Entry Datasheet
| No. | Test | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | Indole | ||||||||||
| 2 | Methyl red | ||||||||||
| 3 | Voges Proskauer's | ||||||||||
| 4 | Citrate utilization | ||||||||||
| 5 | Glucose | ||||||||||
| 6 | Adonitol | ||||||||||
| 7 | Arabinose | ||||||||||
| 8 | Lactose | ||||||||||
| 9 | Sorbitol | ||||||||||
| 10 | Mannitol | ||||||||||
| 11 | Rhamnose | ||||||||||
| 12 | Sucrose |
KB001: HIMVIC™ Biochemical Test Kit (used for differentiation of Enterobacteriaceae species)
Identification Index of various Enterobacteriaceae species
| Test | Indole | Methyl red | Voges Proskauer's | Citrate Utilization | Glucose | Adonitol | Arabinose | Lactose | Sorbitol | Mannitol | Rhamnose | Sucrose |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Budvicia aquatica | + | + | V | + | + | – | V | V | + | + | – | + |
| Buttiauxella agrestis | + | + | V | + | + | + | + | + | + | + | + | + |
| Cedecea davisae | – | + | V | – | + | – | – | V | – | + | – | + |
| Cedecea lapagei | – | + | V | + | + | – | – | V | – | + | – | + |
| Cedecea neteri | – | + | V | – | + | – | – | V | – | + | – | + |
| Citrobacter amalonaticus | – | + | – | V | + | + | + | + | + | + | + | V |
| Citrobacter diversus | + | + | – | V | + | + | + | + | + | + | + | V |
| Citrobacter freundii | – | + | – | + | + | + | + | + | + | + | + | V |
| Enterobacter aerogenes | – | – | + | + | + | + | + | + | + | + | + | + |
| Enterobacter amnigenus (Biogroup I) | – | – | + | V | + | – | + | V | + | + | + | + |
| Enterobacter amnigenus (Biogroup II) | – | V | + | + | + | – | + | V | V | + | + | + |
| Enterobacter taylorae (E. cancerogenus) | – | – | + | + | + | – | – | – | + | + | + | + |
| Enterobacter cloacae | – | – | + | + | + | – | + | + | + | + | + | + |
| Enterobacter gergoviae | + | – | + | + | + | – | + | – | V | + | – | + |
| Enterobacter sakazakii | V | – | + | + | + | + | + | + | + | + | + | + |
| Escherichia coli | + | + | – | – | + | – | – | + | + | + | + | + |
| Escherichia coli, inactive | V | + | – | – | + | – | – | + | + | + | + | + |
| Escherichia blattae | – | + | – | V | + | – | – | – | – | + | V | + |
| Escherichia fergusonii | + | + | – | V | + | – | – | + | + | + | + | V |
| Escherichia hermanii | + | + | – | – | + | – | – | – | V | + | V | V |
| Escherichia vulneris | + | + | – | – | + | – | – | V | + | + | + | – |
| Ewingella americana | – | V | – | + | + | – | – | – | – | + | – | – |
| Hafnia alvei | – | V | V | – | + | – | + | – | – | V | + | – |
| Klebsiella oxytoca | + | – | V | + | + | + | + | + | + | + | + | V |
| Klebsiella pneumoniae subspecies ozaenae | – | – | + | V | + | – | – | V | V | + | V | V |
| Klebsiella pneumoniae subspecies pneumoniae | V | – | + | + | + | + | + | + | + | + | + | + |
| Klebsiella pneumoniae subspecies rhinoscleromatis | – | – | + | – | + | + | – | + | – | + | – | – |
| Klebsiella terrigena | – | V | + | V | + | + | + | + | + | + | + | + |
| Kluyvera ascorbata | + | V | – | + | + | – | + | – | V | + | V | + |
| Leclercia adecarboxylata (Escherichia adecarboxylata) | + | + | – | – | + | – | + | + | – | + | + | V |
| Morganella morganii | + | + | – | – | + | – | – | – | – | – | – | – |
| Pantoea agglomerans | – | V | + | + | + | – | + | V | – | + | + | + |
| Pantoea dispersa | – | V | + | V | + | – | V | – | – | + | – | + |
| Proteus mirabilis | – | + | + | – | + | – | – | – | – | – | – | + |
| Proteus myxofaciens | – | + | + | + | + | – | – | – | – | – | – | – |
| Proteus penneri | – | + | – | – | + | – | – | – | – | – | – | – |
| Proteus vulgaris | + | + | – | V | + | – | – | – | – | – | – | – |
| Providencia alcalifaciens | + | + | – | – | + | – | – | – | – | – | – | V |
| Providencia rettgeri | + | + | – | – | + | – | – | – | – | + | V | V |
| Providencia rustigianii | + | V | – | V | + | – | – | – | – | – | – | V |
| Rahnella aquatilis | – | V | + | V | + | – | + | + | – | + | – | + |
| Salmonella bongori | + | + | – | + | + | – | – | – | + | + | + | + |
| Salmonella choleraesuis subspecies Arizonae | + | + | – | + | + | – | + | V | + | + | + | – |
| Salmonella choleraesuis subsp Choleraesuis | + | + | – | + | + | – | – | – | + | + | + | + |
| Salmonella choleraesuis subspecies Diarizonae | + | + | – | + | + | – | + | V | + | + | + | – |
| Salmonella choleraesuis subspecies Houtenae | + | + | – | + | + | – | – | – | + | + | + | + |
| Salmonella choleraesuis subspecies Indica | + | + | – | + | + | – | + | V | + | + | + | – |
| Salmonella choleraesuis subspecies Salamae | + | + | – | + | + | – | – | – | + | + | + | + |
| Salmonella Enteritidis | + | + | – | + | + | – | – | – | + | + | + | + |
| Salmonella Typhi | – | + | – | – | + | – | – | – | + | + | – | – |
| Serratia entomophila | – | V | V | + | + | + | + | + | – | + | + | + |
| Serratia ficaria | – | V | V | + | + | + | – | V | + | V | + | V |
| Serratia fonticola | + | + | – | + | + | V | V | V | V | + | V | + |
| Serratia marcescens | V | V | V | V | + | – | V | V | V | + | V | V |
| Serratia odorifera (Biogroup I) | V | – | V | + | + | – | V | + | + | + | – | – |
| Serratia odorifera (Biogroup II) | V | V | V | – | + | – | V | V | V | + | V | – |
| Serratia plymuthica | – | V | V | V | + | – | + | V | V | + | + | V |
| Serratia proteamaculans | – | V | V | + | + | + | – | – | V | + | V | + |
| Serratia rubidaea | V | V | V | – | + | – | – | – | – | + | – | – |
| Shigella boydii, Shigella flexneri, Shigella dysenteriae | V | + | – | – | + | – | – | – | V | + | – | – |
| Shigella sonnei | – | – | – | – | + | – | – | – | V | + | V | – |
| Yersinia enterocolitica | V | + | – | + | + | – | + | – | + | + | – | + |
| Yersinia frederiksenii | – | + | V | V | + | – | + | + | + | + | + | + |
| Yersinia intermedia | + | + | V | – | + | – | – | V | V | + | – | + |
| Yersinia pestis | – | V | – | – | + | – | – | – | – | + | – | – |
| Yersinia pseudotuberculosis | – | + | V | – | + | – | – | V | V | + | V | – |
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
- Nd = No data available.
Storage and Shelf Life
On receipt store between 2-8°C. Use before expiry date on the label. Product performance is best if used within stated expiry period.
Disposal
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. 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 (2,3).
| Product Name | HiIMViC™ Biochemical Test Kit |
|---|---|
| SKU | KB001 |
| Customized Product Available | No |
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