Plot No. C40, Road No. 21Y, MIDC, Wagle Industrial Estate,
Thane(W)-400604, MH, India.
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Intended use
Recommended for the susceptibility testing of Haemophilus influenzae.
Composition
| Ingredients | g/L |
|---|---|
| HM infusion B from 300g # | 2.000 |
| Acicase ## | 17.500 |
| Yeast extract | 5.000 |
| Starch | 1.500 |
| Agar | 17.000 |
| Haematin Growth Supplement (FD117) | 1 vial |
| NAD (2x7.50 mg) | 15.000 mg |
| Haematin (2x7.50 mg) | 15.000mg |
| Final pH (at 25°C) | 7.4±0.2 |
**Formula adjusted, standardized to suit performance parameters
# - Equivalent to Beef infusion from, ##-Equivalent to casein acid hydrolysate
Directions
Either streak, inoculate or surface spread the test inoculum (50-100 cfu) aseptically on the plate.
Principle And Interpretation
Haemophilus species are nutritionally fastidious in nature. They require either exogenous hemin (X-factor) or NAD (V-factor) or both (1). Due to this reason, Mueller Hinton Agar, which is used for antimicrobial susceptibility of bacteria (2,3,4), can't be used for the antimicrobial susceptibility testing of Haemophilus. Haemophilus Test Agar Base, studied by Jorgensen et al (5,6) is used for the susceptibility testing of Haemophilus influenzae. This medium has similar composition as Mueller Hinton Agar, with the addition of yeast extract and added growth supplements. Haemophilus Test Agar Base is simple, transparent and posses minimum risk of antagonism of antimicrobial agents (5). Haemophilus Test Agar Base is also recommended by the United States National Committee for Clinical Laboratory Standards (NCCLS) for both dilution and disc diffusion assays (7). This medium scores over Mueller Hinton Agar with heamoglobin over clarity, thereby enabling proper visualization of inhibition zones. It also has low levels of the nucleotide thymidine, which allows testing of trimethoprim / sulphamethoxazole.
Haemophilus Test Agar Base contains HM infusion B from and acicase, which provide essential nutrients to the organisms. Yeast extract serves as a source of B complex vitamins. Starch acts as a protective colloid against toxic substances present in the medium. The surface of a Haemophilus Test Agar Base with added nutrients is inoculated either by using swab or by spreading the suspension. Antimicrobial discs i.e. paper discs impregnated with specific amount of antibiotics or other antimicrobial agents are placed on the surface of medium spaced properly. The plates are incubated in a CO2 incubator and subsequently the inhibition zones around each disc are read. Comparing the zones of inhibition with the NCCLS standards, the determination as to whether the organism is susceptible, resistant or intermediate in its response to the antimicrobial substances is made (7).
Type of specimen
Isolated Microorganism from clinical samples.
Specimen Collection and Handling:
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (3,5).
After use, contaminated materials must be sterilized by autoclaving before discarding.
Warning and Precautions :
In Vitro diagnostic Use only. For professional use only. Read the label before opening the container. 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. Safety guidelines may be referred in individual safety data sheets.
Limitations :
- The plates are to be incubated at 5-7% carbon dioxide at 35-37°C for 18-24 hours for appropriate results.
- Individual organisms differ in their growth requirement and may show variable growth patterns on the medium.
- Each lot of the medium has been tested for the organisms specified on the COA. It is recommended to users to validate the medium for any specific microorganism other than mentioned in the COA based on the user's unique requirement.
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 Haemophilus Test Agar in 90mm disposable plate with smooth surface and absence of black particles/cracks/bubbles
Colour of medium
Light amber coloured medium
Quantity of medium
25 ml of medium in 90 mm plate
pH
6.80 -7.20
Sterility Check
Passes release criteria
Cultural Response
Cultural characteristics observed in 5-7% carbon dioxide after an incubation at 35-37°C for 18-24 hours.
| Organism | Inoculum (CFU) | Growth | Recovery |
|---|---|---|---|
| Haemophilus influenzae ATCC 49766 | 50-100 | luxuriant | >=70% |
| Enterococcus faecalis ATCC 29212 (00187*) | 50-100 | good-luxuriant | >=70% |
| Streptococcus pyogenes ATCC 19615 | 50-100 | good-luxuriant | >=70% |
| Neisseria meningitidis ATCC 13090 | 50-100 | good-luxuriant | >=70% |
| Staphylococcus aureus subsp.aureus ATCC 25923 (00034*) | 50-100 | good-luxuriant | >=70% |
(*) - Corresponding WDCM numbers
Storage and Shelf Life
Store between 2-8°C. Use before expiry period on the label. Product performance is best if used within stated expiry period.
Disposal
User must ensure safe disposal by autoclaving and/or incineration of used or unusable preparations of this product. 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 (3,5).
| Product Name | Haemophilus Test Agar Plate |
|---|---|
| SKU | MP1259 |
| Customized Product Available | No |
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