HiCrome™ UTI HiCynth™ Agar

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MCD1353
Recommended for presumptive identification and confirmation of microorganisms mainly causing urinary tract infections, can also be used for testing water, food, environmental and other clinical samples.


Intended use

Recommended for presumptive identification and confirmation of microorganisms mainly causing urinary tract infections, can also be used for testing water, food, environmental and other clinical samples.

Composition**

Ingredients g/L
HiCynth™ Peptone No.1* 15.000
Chromogenic mixture 2.450
Agar 15.000
Final pH (at 25°C) 6.8±0.2

**Formula adjusted, standardized to suit performance parameters
*Chemically defined peptone

Directions

Suspend 32.45 gram in 1000 ml purified / distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Cool to 45-50°C. Mix well and pour into sterile Petri plates.

Principle And Interpretation

Urinary tract infections are bacterial infections affecting parts of urinary tract. The common symptoms of urinary tract infection are urgency and frequency of micturition, with associated discomfort or pain. The common condition is cystitis, due to infection of the bladder with an uropathogenic bacterium, which most frequently is Escherichia coli, but sometimes Staphylococcus saprophyticus or especially in hospital-acquired infections, Klebsiella species, Proteus mirabilis, other coliforms, Pseudomonas aeruginosa or Enterococcus faecalis (1). HiCrome™ UTI Agar is formulated on basis of work carried out by Pezzlo (2) Wilkie et al (3), Friedman et al (4), Murray et al (5), Soriano and Ponte (6) and Merlino et al (7). HiCrome™ UTI HiCynth™ Agar is modification of the same, prepared by completely replacing animal or vegetable based peptones with chemically defined peptone.

It facilitates and expedites the identification of some gram-negative bacteria and some gram-positive bacteria on the basis of different contrasted colony colours produced by reactions of genus or species specific enzymes with two chromogenic substrates. The chromogenic substrates are specifically cleaved by enzymes produced by Enterococcus species, E.coli and coliforms. Presence of amino acids like phenylalanine and tryptophan from peptones helps for detection of tryptophan deaminase activity, indicating the presence of Proteus species, Morganella species and Providencia species.

One of the chromogenic substrate is cleaved by β-glucosidase possessed by Enterococci resulting in formation of blue colonies. E.coli produce pink colonies due to the enzyme β-D-galactosidase that cleaves the other chromogenic substrate. Further confirmation of E.coli can be done by performing the indole test. Coliforms produce purple coloured colonies due to cleavage of both the chromogenic substrate. Colonies of Proteus, Morganella and Providencia species appear brown because of tryptophan deaminase activity. HiCynth™ Peptone No.1 provides nitrogenous, carbonaceous compounds and other essential growth nutrients. This medium can be made selective by supplementation with antibiotics for detecting microorganisms associated with hospital borne infections.

Type of specimen

Clinical samples: urine, faeces; Food samples; Water samples

Specimen Collection and Handling

For clinical samples follow appropriate techniques for handling specimens as per established guidelines (8,9).

For food samples, follow appropriate techniques for sample collection and processing as per guidelines (10).

For water samples, follow appropriate techniques for sample collection, processing as per guidelines (11).

After use, contaminated materials must be sterilized by autoclaving before discarding.

Warning and Precautions

In Vitro diagnostic use. 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

  1. Since it is an enzyme-substrate based reaction, the intensity of colour may vary with isolates.
  2. Some species may show variable growth due to difference in nutritional requirements.

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
Cream to yellow homogeneous free flowing powder

Gelling
Firm, comparable with 1.5% Agar gel

Colour and Clarity of prepared medium
Light amber coloured, clear to slightly opalescent gel forms in Petri plates

Reaction
Reaction of 3.24% w/v aqueous solution at 25°C. pH : 6.8±0.2

pH
6.60-7.00

Cultural Response
Cultural characteristics observed after an incubation at 35-37°C for 16-24 hours.

Organism Inoculum (CFU) Growth Recovery Colour of Colony
Enterococcus faecalis ATCC 29212 (00087*) 50-100 luxuriant >=70% blue, small
Escherichia coli ATCC 25922 (00013*) 50-100 luxuriant >=70% pink-purple
Klebsiella pneumoniae ATCC 13883 (00097*) 50-100 luxuriant >=70% blue to purple, mucoid
Pseudomonas aeruginosa ATCC 27853 (00025*) 50-100 luxuriant >=70% colourless (greenish pigment may be observed)
Proteus mirabilis ATCC 12453 50-100 luxuriant >=70% light brown
Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) 50-100 luxuriant >=70% golden yellow

Key: (*) Corresponding WDCM numbers.

Storage and Shelf Life

Store between 15-25°C in a tightly closed container and the prepared medium at 2-8°C. Use before expiry date on the label. On opening, product should be properly stored dry, after tightly capping the bottle in order to prevent lump formation due to the hygroscopic nature of the product. Improper storage of the product may lead to lump formation. Store in dry ventilated area protected from extremes of temperature and sources of ignition Seal the container tightly after use. 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 (8,9).

More Information
Product Name HiCrome™ UTI HiCynth™ Agar
SKU MCD1353
Product Type HiCynth™, HiCrome™
Physical Form Powder
Origin Chemically defined (HiCynth™)
Packaging type HDPE
References 1.Collee J. G., Fraser A. G., Marmion B. P., Simmons A., (Eds.), Mackie and McCartney, Practical Medical Microbiology,1996, 14th Edition, Churchill Livingstone.2.Pezzlo M., 1998, Clin. Microbiol. Rev., 1:268-280.3.Wilkie M. E., Almond M. K., Marsh F. P., 1992, British Medical Journal 305:1137-1141.4.Friedman M. P. et al, 1991, J. Clin. Microbiol., 29:2385-2389.5.Murray P., Traynor P. Hopson D., 1992, J. Clin. Microbiol., 30:1600- 1601.6.Soriano F., Ponte C., 1992, J. Clin. Microbiol., 30:3033-3034.7.Merlino et al, 1995, Abstr. Austr. Microbiol. 16(4):17-3.
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