HiCombi™ VRBGA - VRBGA Agar Plate

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HB015


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Intended Use

Recommended for the selection and subculture of bile tolerant organisms in accordance with the harmonized method of USP/EP/BP/JP/IP.

Composition

Ingredients Gms / Litre
Yeast extract 3.000
Gelatin peptone # 7.000
Bile salts 1.500
Sodium chloride 5.000
Glucose monohydrate 10.000
Agar 15.000
Neutral red 0.030
Crystal violet 0.002
pH after heating (at 25°C) 7.4±0.2

**Formula adjusted, standardized to suit performance parameters

# Pancreatic digest of gelatin

Directions

Streak the test inoculum (50-100 CFU) aseptically.

Principle and Interpretation

Violet Red Bile Glucose Agar is a selective medium recommended for detection and enumeration of Enterobacteriaceae especially the bile tolerant gram negative bacteria in accordance with the microbial limit testing by harmonized methodology of USP/EP/BP/JP/IP (7,1,2,3,4) from non-sterile products and pharmaceutical preparations. Gelatin peptone and yeast extract provide nitrogenous, carbonaceous compounds, long chain amino acids, vitamins and other nutrients essential for bacterial metabolism. This media is selective due to presence of the inhibitors; bile salts positive organisms especially Staphylococci. Neutral red indicator helps to detect glucose fermentation. Glucose is the fermentable carbohydrate, utilization of which leads to the production of acids. Neutral red indicator detects the acidity so formed. Crystal violet and bile salts mixture help to inhibit the accompanying gram-positive and unrelated flora. Sodium chloride maintains the osmotic equilibrium. Further biochemical tests are necessary for positive identification (7).

Type of Specimen

Pharmaceutical samples

Specimen Collection and Handling

For pharmaceutical samples, follow appropriate techniques for sample collection, processing as per guidelines (8,1,2,3,4). After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions

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 specimens. Safety guidelines may be referred in individual safety data sheets.

Limitations

  • 1. Though the medium is for selective isolation of Enterobacteriaceae, further biochemical and serological testing must be carried out for further confirmation.
  • 2. Over incubation may result in reverting of reaction.

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 VRBGA - VRBGA Agar in 90 mm disposable plate.

Colour: Red with purplish tinge coloured medium

Quantity of Medium: 10 ml of medium in each side of 90mm disposable Biplate.

pH: 7.20 -7.60

Sterility Test: Passes release criteria

Cultural Response

Growth Promotion is carried out in accordance with the harmonized method of USP/EP/BP/JP. Cultural response was observed after incubation at 30-35°C for 18-24 hours. Recovery rate is considered as 100% for bacteria growth on Soyabean Casein Digest Agar.

Growth Promoting Properties

Growth of microorganism comparable to that previously obtained with previously tested and approved lot of medium occurs at the specified temperature for not more than the shortest period of time specified inoculating <=100 cfu (at 30-35°C for <=18 hours).

Indicative Properties

Colonies are comparable in appearance and indication reaction to those previously obtained with previously tested and approved lot of medium occurs for the specified temperature for a period of time within the range specified inoculating 100 cfu (at 30-35°C for 18-24hours).

Organism Inoculum (CFU) Growth Observed Lot value (CFU) Recovery Colour of colony Incubation temperature
Growth Promoting + Indicative
Escherichia coli ATCC 8739 (00012*) 50-100 luxuriant 25-100 >=50% pink-red with bile precipitate 18-24 hrs
Pseudomonas aeruginosa ATCC 9027 (00026*) 50-100 luxuriant 25-100 >=50% pink to red 18-24 hrs
Additional Microbiological Testing
Escherichia coli NCTC 9002 50-100 good-luxuriant 25-100 >=50% pink-red with bile precipitate 18-24 hrs
Escherichia coli ATCC 25922 (00013*) 50-100 good-luxuriant 25-100 >=50% pink-red with bile precipitate 18-24 hrs
Salmonella Enteritidis ATCC 13076 (00030*) 50-100 good-luxuriant 25-100 >=50% light pink 18-24 hrs
# Klebsiella aerogenes ATCC 13048 (00175*) 50-100 good-luxuriant 25-100 >=50% pink-red 18-24 hrs
Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) >=103 inhibited 0 0% >=24 hrs
Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) >=103 inhibited 0 0% >=24 hrs

Key :- (#) Formerly known as Enterobacter aerogenes (*) Corresponding WDCM numbers

Storage and Shelf Life

Store between 20-30°C. Use before expiry date 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 sample must be decontaminated and disposed of in accordance with current laboratory techniques (5,6).

More Information
Product Name HiCombi™ VRBGA - VRBGA Agar Plate
SKU HB015
Customized Product Available No
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