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HiCarbo™ Kit

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SKU:
KB009

Intended use

KB009 is a combination of 35 tests for utilization of carbohydrate tests from clinical specimen and non clinical samples. It can also be used for validating known laboratory strains.

Note: KB009 is available as a total kit and also individually Part A, Part B and Part C

Kit contents

  1. Each kit contains 5/10/20 kits of Part A, Part B and Part C of KB009, sufficient material to perform 5/10/20 tests.
  2. Part A: HiCarbo™ Kit- Part A: Lactose, Xylose, Maltose, Fructose, Dextrose, Galactose, Raffinose, Trehalose, Melibiose, Sucrose, L- Arabinose, Mannose.
  3. Part B: HiCarbo™ Kit- Part B: Inulin, Sodium gluconate, Glycerol, Salicin, Dulcitol, Inositol, Sorbitol, Mannitol, Adonitol, Arabitol, Erythritol, a-Methyl-D-glucoside.
  4. Part C : HiCarbo™ Kit- Part C: Rhamnose, Cellobiose, Melezitose, a-Methyl-D-Mannoside, Xylitol, ONPG, Esculin, D- Arabinose, Citrate, Malonate, Sorbose and 1 control.
  5. Technical product insert.
  6. Result Interpretation Chart and Result Entry Datasheet.

Material Required but not supplied

  1. McFarland standard
  2. Inoculation loops, pipettes
  3. Enrichment medium / Isolation media

Direction

Preparation of inoculum :

  • KB009 cannot be used directly on clinical specimens. The organisms to be identified have to be first isolated and purified. Only pure cultures should be used. Isolate the organism to be identified on a common medium like Nutrient Agar (M001/ Nutrient Agar for Oxidase (M1274) or BHI Agar (M211).
  • 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 0.5 OD at 620nm. Some fastidious organisms may require more than 6 hours of incubation. In this case incubate till the inoculum turbidity reaches 0.5 OD at 620nm.
  • Alternatively, prepare the inoculum by picking 1-3 well isolated colonies and make a homogeneous suspension in 2-3ml sterile saline. The density of the suspension should be 0.5 OD at 620nm.

Inoculation of the kit:

  1. Open the kit aseptically. Peel off the sealing foil.
  2. Inoculate each well with 50 µl of the above inoculum by surface inoculation method.
  3. Alternatively, the kit can also be inoculated by stabbing each individual well with a loopful of inoculum.

Incubation:

Temperature of incubation: 35±2°C. Duration of incubation: 18-24 hours

Interpretation of results :

Interpret results as per the result interpretation chart.

Principle

Each HiCarbo™ kit is a standardized calorimetric identification system utilizing thirty five 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 spontaneous colour change in the media.

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

  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 upto 48 hours. Orange colour after 48 hours of incubation should be interpreted as a negative reaction.
  3. At times organisms give conflicting result because of mutation or the media used for isolation, cultivation and maintenance.
  4. The identification index has been compiled from standard references and results of tests carried out in the laboratory.
  5. Erroneous false negative results may be obtained if the inoculum turbidity is less than 0.5 OD.
  6. Results are more prominent if an enriched culture is used instead of a suspension.
  7. 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 of Part A, Part B each having 12 carbohydrates utilization tests and Part C containing 11 tests and 1 control.

Part A:

Lactose, Xylose, Maltose, Fructose, Dextrose, Galactose, Raffinose, Trehalose, Melibiose, Sucrose, L-Arabinose, Mannose.

Part B:

Inulin, Sodium gluconate, Glycerol, Salicin, Dulcitol, Inositol, Sorbitol, Mannitol, Adonitol, Arabitol, Erythritol, a-Methyl-D-glucoside.

Part C:

Rhamnose, Cellobiose, Melezitose, a-Methyl-D-Mannoside, Xylitol, ONPG, Esculin, D-Arabinose, Citrate, Malonate, Sorbose and1 control.

Quantity of medium

0.8 ml of medium in each well.

Sterility Check

Passes release criteria

Interpretation of results :

Interpret results as per the result interpretation chart.

  1. Carbohydrate utilization Test : Well No. 1-12 of Part A, B and Well No. 1-5, 8 and 11 of C
    • Positive test is indicated by a colour change to yellow colour.
    • Red or no colour change indicates a negative reaction.
  2. ONPG Test:Well No. 6 of Part C
    • Medium changes from colourless to yellow if the test is positive.
    • Medium remains colourless if the test is negative.
  3. Esculin Hydrolysis: Well No. 7 of Part C
    • Colour of the medium changes from cream to black if the test is positive.
    • Medium remains cream in colour if the test is negative.
  4. Citrate Utilization: Well No. 9 of Part C
    • Colour of the medium changes from green to blue if the test is positive.
    • Medium remains green in colour if the test is negative.
  5. Malonate Utilization: Well No. 10 of Part C
    • Colour of the medium changes from light green to blue if the test is positive.
    • Medium remains light green in colour if the test is negative.

Result Interpretation chart

Test Principle Original colour of the medium Positive reaction Negative reaction
Carbohydrate utilization Detects carbohydrate utilization Pinkish Red / Red Yellow Red / Pink
ONPG Detects β-galactosidase activity Colourless Yellow Colourless
Esculin hydrolysis Detects esculin hydrolysis Cream Black Cream
Citrate utilization Detects capability of organism to utilize citrate as a sole carbon source Green Blue Green
Malonate utilization Detects capability of organism to utilize sodium malonate as a sole carbon source Light green Blue Light green

Result Entry Datasheet

The kit includes the following 35 tests:

  1. Lactose
  2. Xylose
  3. Maltose
  4. Fructose
  5. Dextrose
  6. Galactose
  7. Raffinose
  8. Trehalose
  9. Melibiose
  10. Sucrose
  11. L-Arabinose
  12. Mannose
  13. Inulin
  14. Sodium gluconate
  15. Glycerol
  16. Salicin
  17. Dulcitol
  18. Inositol
  19. Sorbitol
  20. Mannitol
  21. Adonitol
  22. Arabitol
  23. Erythritol
  24. a-Methyl-D-glucoside
  25. Rhamnose
  26. Cellobiose
  27. Melezitose
  28. a-Methyl-D-mannoside
  29. Xylitol
  30. ONPG
  31. Esculin hydrolysis
  32. D-Arabinose
  33. Citrate utilization
  34. Malonate utilization
  35. Sorbose

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).

More Information
Product Name HiCarbo™ Kit
SKU KB009
Customized Product Available No
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