HiAcinetobacter™ Identification Kit

Product Identifier

Introduction

KB014 is identification system for Acinetobacter comprising of twenty four biochemical tests (Twelve tests in Part A and Twelve tests in part B). The tests include the IMVIC group of tests, carbohydrate fermentation tests, amino acid hydrolysis tests and other tests. The complete list of organisms that can be identified with this system is given in the identification index provided with the kit.

Principle

KB014 is a standardized, colorimetric identification system utilizing twenty four 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.

Kit Contents

Instructions for use

Note: KB014 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.

Preparation of inoculum

• Isolate the organism to be identified on a common medium like Nutrient Agar (M001) or Soyabean Casein Digest Agar (M290). Pick up a single isolated colony and inoculate in 5 ml Brain Heart Infusion Broth and incubate at 37°C for 4-6 hours until the inoculum turbidity is 0.10D at 620nm or 0.5 McFarland standards. Some fastidious organisms may require more than 6 hours of incubation. In this case incubate till the inoculum turbidity reaches 0.10D 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 0.10D at 620nm.

Note: Erroneous false negative results may be obtained if the inoculum turbidity is less than 0.1 OD.Results are more prominent if an enriched culture is used instead of suspension.

Inoculation of the strip:

• Open the kit aseptically.
• Inoculate each well with 50 µl of the above inoculum by surface inoculation method.
• Alternatively, the strip can 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 result interpretation chart. Addition of reagents in well no 1, 2, 7 & 10 of part A should be done at the end of incubation period that is after 18 - 24 hours.

Biochemical Reactions (Part A)

Carbohydrate Fermentation Tests (Part B)

Important Considerations for Interpretation

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. At times organisms give conflicting result because of mutation or the media used for isolation, cultivation & maintenance.
4. The identification index has been compiled from standard references & 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 & handling of the strips.
• Reagents should not come in contact with skin, eyes or clothing.

Disposal

After use, strips and the instruments used for isolation & inoculation (pipettes, loops etc.) must be disinfected using a suitable disinfectant and then discarded by incineration or autoclaving in a disposal bag.

Storage and Shelf-life