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HiCombi™ Dual Performance Salmonella Medium - XLD
Intended use
A qualitative test for rapid growth and confirmation of Salmonella. Combination of solid (7 ml) media and liquid (20 ml) media in single bottle.
Composition
| Ingredients | g/L |
|---|---|
| 7 ml Solid Phase | |
| Yeast extract | 3.000 |
| L-Lysine | 5.000 |
| Lactose | 7.500 |
| Sucrose | 7.500 |
| Xylose | 3.500 |
| Sodium chloride | 5.000 |
| Sodium deoxycholate | 2.500 |
| Sodium thiosulphate | 6.800 |
| Ferric ammonium citrate | 0.800 |
| Phenol red | 0.080 |
| Agar | 15.000 |
| 20 ml Liquid Phase | |
| Yeast extract | 3.000 |
| L-Lysine | 5.000 |
| Lactose | 7.500 |
| Sucrose | 7.500 |
| Xylose | 3.500 |
| Sodium chloride | 5.000 |
| Sodium deoxycholate | 2.500 |
| Sodium thiosulphate | 6.800 |
| Ferric ammonium citrate | 0.800 |
| Phenol red | 0.080 |
| Final pH (at 25°C) | 7.4±0.2 |
**Formula adjusted, standardized to suit performance parameters
Directions
Label the ready to use HiCombi T™ Dual performance medium bottle. Do not unscrew the cap. Remove the top seal of the cap. Disinfect the part of the rubber stopper which is now exposed. Transfer the Sample into the culture bottle by puncturing the rubber stopper with the sterile, disposable needle and syringe. Venting may be carried for aerobic cultures as in Hisafe Dual performance culturing system. Incubate the bottle for 4-6 hours at 30 -35°C. For adsorption on solid surface. Tilt the bottle horizontally whereby solid medium is submerged with liquid phase. DO NOT SHAKE OR HOLD MORE THAN 15 SECONDS. Revert into an upright position and incubate for 18-24 hours at 30-35°C or longer if necessary. Venting: Use sterile venting needle (LA038). Keep the bottle in an upright position preferably in a biological safety cabinet, place an alcohol swab over the rubber stopper and insert the venting needle with filter through it. Insertion and withdrawal of the needle should be done in a straight line. Discard the needle and mix the contents by gently inverting the bottle 2-3 times. Do not vent the bottle for anaerobic cultures. Recommended volume of blood to be tested in LQ030: 3-5 ml (For Paediatric use).
Principle And Interpretation
Hicombi™ Dual performance Medium contains in a single glass bottle, a combination of 20 ml of broth and one Agar coated surface. Both media are rich in growth factors. The Hicombi™ Dual Culturing system is a fast, efficient and simpler detection and preliminary identification system for microorganism. Two in one step procedure involving inoculation and simultaneous isolation on solid surface. XLD Agar has been recommended for the identification of Enterobacteriaceae (1). It is both selective and differential medium. The medium contains yeast extract, which provides nitrogen and vitamins required for growth. Though the sugars xylose, lactose and sucrose provide sources of fermentable carbohydrates, xylose is mainly incorporated into the medium since it is not fermented by Shigellae but practically by all enterics. This helps in the differentiation of Shigella species. Sodium chloride maintains the osmotic balance of the medium. Lysine is included to differentiate the Salmonella group from the non-pathogens. Salmonellae rapidly ferment xylose and exhaust the supply. Subsequently lysine is decarboxylate by the enzyme lysine decarboxylase to form amines with reversion to an alkaline pH that mimics the Shigella reaction. However, to prevent this reaction by lysine-positive coliforms, lactose and sucrose are added to produce acid in excess. Degradation of xylose, lactose and sucrose to acid causes phenol red indicator to change its colour to yellow. Bacteria that decarboxylate lysine to cadaverine can be recognized by the appearance of a red colouration around the colonies due to an increase in pH. These reactions can proceed simultaneously or successively, and this may cause the pH indicator to exhibit various shades of colour or it may change its colour from yellow to red on prolonged incubation. To add to the differentiating ability of the formulation, an H2S indicator system, consisting of sodium thiosulphate and ferric ammonium citrate, is included for the visualization of hydrogen sulphide produced, resulting in the formation of colonies with black centers. The non-pathogenic H2S producers do not decarboxylase lysine; therefore, the acid reaction produced by them prevents the blackening of the colonies (2).
Type of specimen
Clinical samples - Blood
Specimen Collection and Handling
For clinical samples follow appropriate techniques for handling specimens as per established guidelines (3,4).
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
- Slight precipitation in the medium may occur, which is inheritant property of the medium,and does not affect the performance of the medium.
- Some species may show poor growth due to nutritional variations.
- Some Proteus strains may give red to yellow colouration with most colonies developing black centers, giving rise to false positive reactions.
- Non-enterics like Pseudomonas and Providencia may exhibit red colonies.
- S. Paratyphi A, S.Choleraesuis, S.Pullorum and S.Gallinarum may form red colonies without H2S, thus resembling Shigella species.
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: In a sterile glass bottle combination of broth and one agar coated surface.
Colour of Agar medium: Red coloured Agar media
Colour of liquid medium: Red coloured liquid medium
Quantity of medium: 7 ml of solid media in glass bottle and 20 ml of liquid medium in glass bottle
pH of Agar medium: 7.20-7.60
pH of liquid medium: 7.20-7.60
Sterility Check: Passes release criteria
Cultural response: Cultural characteristics was observed after incubation at 35-37°C for 18-72 hours.
Cultural Characteristics
| Organism | Inoculum (CFU) | Growth in liquid medium | Growth on agar medium | Colour of colony |
|---|---|---|---|---|
| Salmonella Typhimurium ATCC 14028 (00031*) | 50-100 | luxuriant | good-luxuriant | red with black |
| Salmonella Abony NCTC 6017 (00029*) | 50-100 | luxuriant | good-luxuriant | red with black centres |
| Escherichia coli ATCC 8739 (00012*) | 50-100 | fair | fair | yellow |
| Escherichia coli ATCC 25922 (00013*) | 50-100 | fair | fair | yellow |
| ## Proteus hauseri ATCC 13315 | 50-100 | luxuriant | good-luxuriant | grey with black centres |
| Salmonella Paratyphi A ATCC 9150 | 50-100 | luxuriant | good-luxuriant | red |
| Salmonella Paratyphi B ATCC 8759 | 50-100 | luxuriant | good-luxuriant | red with black centres |
| Salmonella Enteritidis ATCC 13076 (00030*) | 50-100 | luxuriant | good-luxuriant | red with black centres |
| Salmonella Typhi ATCC 6539 | 50-100 | luxuriant | good-luxuriant | red with black centres |
| Shigella dysenteriae ATCC 13313 | 50-100 | luxuriant | good-luxuriant | red |
| Shigella flexneri ATCC 12022 (00126*) | 50-100 | luxuriant | fair-good | red |
| Shigella sonnei ATCC 25931 | 50-100 | luxuriant | fair-good | red |
| # Klebsiella aerogenes ATCC 13048 (00175*) | 50-100 | fair | fair | yellow |
| Enterobacter cloacae ATCC 13047 (00083*) | 50-100 | luxuriant | fair | yellow |
| Staphylococcus aureus subsp. aureus ATCC 25923 (00034*) | >=104 | inhibited | inhibited | |
| Staphylococcus aureus subsp. aureus ATCC 6538 (00032*) | >=104 | inhibited | inhibited | |
| Enterococcus faecalis ATCC 29212 (00087*) | >=104 | inhibited | inhibited |
Key: (*) Corresponding WDCM numbers.
## Formerly known as Proteus vulgaris
# formerly known as Enterobacter aerogenes
Storage and Shelf Life
On receipt store between 2-8°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 clinical sample must be decontaminated and disposed of in accordance with current laboratory techniques (3,4).
| Product Name | HiCombi™ Dual Performance Salmonella Medium - XLD |
|---|---|
| SKU | LQ030 |
| Customized Product Available | No |

