HiBifido™ Identification Kit

Instructions for use Interpretation of Results

Interpret results as per the standards given in the result interpretation chart. Addition of 3% H₂O₂ in well no 1, should be done at the end of incubation period that is after 24-48 hours under anaerobic condtions.

Well No. 1: Catalase Test:

• Scrape a loopful of growth from the surface of the well. Dip the loop in a small, clean test tube containing 3% Hydrogen peroxide.
• Positive catalase test is seen as effervescence coming out from the surface of the loop.
• No effervescence is observed in case of negative catalase test.

Note : 3% H₂O₂ has to be freshly prepared

Important points to be taken into consideration while interpreting the result

• Allow the reagents to come to room temperature after removal from the refrigerator.

Disposal of used material

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

Instructions for use

Preparation of inoculum :

• Isolate the organism to be identified on a medium like Bifidobacterium Agar (M1396) or Soyabean Casein Digest Agar (M290) with addition of 5-7% v/v defibrinated sheep blood.
• Pick up a single isolated colony and inoculate in 5 ml Bifidobacterium Broth (M1395) and incubate at 35-37C under anaerobic condition for 6-18 hours until the inoculum turbidity is 0.1OD at 620nm or 0.5 McFarland standards.
• 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 0.1OD at 620nm or 0.5 McFarland standards.

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 kit :

• Open the kit aseptically. Peel off the sealing tape.
• Inoculate each well with 50 µl of the above inoculum by surface inoculation method.
• Alternatively the kit can be inoculated by stabbing each individual well with a loopful of inoculum.

Incubation :

• Temperature of incubation: 35-37°C under anaerobic condtions. Duration of incubation : 24-48 hours.

Storage and Shelf life :

On receipt, store at 2-8°C. Shelf life is 12 months. Product performance is best if used within stated expiry period.

Biochemical reactions of KB021