GlabrataQuick™ KIT

Cat. no. Z298 GlabrataQuick™ Kit 24 tests/kit
Each kit contains:
8-Well Reaction Strips 24 strips
Color Reaction Reader 1 reader
Inoculation Buffer, 1ml 24 tubes

INTENDED USE

Hardy Diagnostics GlabrataQuick™ Kit is a rapid test for the identification of Candida ( Torulopsis ) glabrata . GlabrataQuick™ Kit uses acid production from three carbohydrates to identify Candida ( Torulopsis ) glabrata in one to two hours.

SUMMARY

In clinical laboratories, Candida albicans is the most frequently isolated species as the cause of disease. However, the need to screen for Candida glabrata has risen in importance as it has been isolated as the cause of major infections, particularly in immunosuppressed patients. Increasing resistance to fluconazole is common. Apart from multifocal, disseminated disease, it is often recovered from urine specimens and has been estimated to account for as many as 21% of urinary yeast isolates.

Hardy Diagnostics GlabrataQuick™ Kit is a rapid test method for the identification of C. glabrata on the basis of rapid trehalose assimilation at 35ºC.

Rapid Trehalose Medium is derived from the formula described by Stockman and Roberts. (6) It utilizes yeast nitrogen base as a source of nitrogenous compounds. The yeast nitrogen base is low in other carbohydrates that might interfere with the rapid test. Bromcresol green is the indicator, allowing the visualization of an acid shift that is indicative of a positive reaction. Trehalose is incorporated into the medium at a high concentration as the carbon source. This product is best used in conjunction with HardyCHROM™ Candida (Cat. no. G301). On this medium, C. glabrata will appear as smooth pink colonies, often with a darker pink or mauve center, and should be confirmed with the trehalose test.

The GlabrataQuick™ Kit reduces the time required to determine acid production to one to two hours. C. glabrata will be positive in the trehalose wells and negative in the maltose and sucrose wells. Rare strains of C. albicans , C. parapsilosis , C. guilliermondii , C. lusitaniae , C. tropicalis , C. krusei and Saccharomyces cerevisiae that might produce a positive reaction in trehalose wells will also produce a positive reaction in maltose or sucrose wells.

FORMULA

Ingredients per liter of deionized water:*

GlabrataQuick™ Medium Base (well A)
Yeast Nitrogen Base 13.4gm
Bromcresol Green 0.8gm
In addition, the following wells are supplemented with a specific carbohydrate:
Trehalose (wells in row C)
Maltose (wells in row D)
Sucrose (wells in row E)




GlabrataQuick™ Inoculation Buffer, 1ml
pH Buffers 13.6gm

Final pH 5.4 +/- 0.1 at 25ºC.

* Adjusted and/or supplemented as required to meet performance criteria.

STORAGE AND SHELF LIFE

Storage: Upon receipt store at 2-8ºC. away from direct light. The kit should not be used if there is any discoloration or if the expiration date has passed. Do not use if desiccant is missing or if lids have been removed. Product is light and temperature sensitive; protect from light, excessive heat, moisture, and freezing.

PRECAUTIONS

PROCEDURE

Specimen Collection: This product is not intended for the primary isolation of patient specimens. This product is intended to be used to identify cultures of isolated organisms.

The appropriate organisms for performing the GlabrataQuick™ test are non-capsulated yeast, germ tube negative or smooth pink colonies often with a darker pink center that have been isolated on the HardyCHROM™ Candida plates (Cat. no. G301).

Method of use:

1. Remove caps from the test well strips to be used and leave the strips in the base. Unused well strips should be removed from the base and stored in the bag (with caps on) until needed.

2. Obtain isolated colonies that are 24-48 hours old, preferably from HardyCHROM™ Candida (Cat. no. G301). Prepare a very heavy suspension using the Inoculation Buffer tubes provided. Vortex suspension for 30 seconds. If multiple specimens are processed at the same time, make sure to vortex again before inoculating the wells, to prevent settling of the yeast cells. A low density inoculum may result in false-negative reactions.

3. Aseptically transfer approximately 0.1ml (or four to five drops from a transfer pipet) of the suspension into wells A, C, D, and E of the strip so that the wells are half full. Do not add any suspension to the second (B) or sixth (F) wells. These wells are intentionally left empty. Do not remove the test well strips from the base holder.

4. Incubate well strips uncovered at 35ºC. aerobically. Do not incubate in a CO 2 atmosphere. The carbohydrate wells (rows A, C, D and E) may be read after one hour. A final reading may be done at two hours. See "Limitations" below.

5. Place the tray with the wells on the "Color Reaction Reader" card, included with the kit, to record the results.

INTERPRETATION OF RESULTS

A change of color from blue or blue-green to yellow or greenish-yellow is considered positive for the carbohydrate wells (rows C, D and E). Well A is used as a reference and should remain negative (blue or blue-green) throughout incubation, since it contains no carbohydrate. Final results must be read no later than three hours after incubation is started.
Note: Color changes in the carbohydrate wells that take place after three hours of incubation time should be disregarded.

Organism Negative
Control
(well A)



Trehalose
(well C)
Maltose
(well D)
Sucrose
(well E)


C. glabrata -
blue / blue-green
+
yellow / greenish-yellow
-
blue / blue-green / green
-
blue / blue-green / green

Non- C. glabrata
-
blue / blue-green


if yellow to greenish-yellow
in well D or E
then isolate is not C. glabrata regardless of
trehalose reaction


* Some strain to strain variation may occur, and isolates may require more biochemical and/or serological tests for complete identification. Refer to the listed references for more information regarding the identification and differentiation of Candida species.

LIMITATIONS

This medium is intended to be used an identification method for C. glabrata . Other biochemical and/or serological tests may be required for complete identification of this organism.

It is recommended to report final results at two hours. If the inoculation time inadvertently extends beyond two hours the results are still valid. The test must be repeated if the incubation time extends beyond three hours.

A very heavy inoculum must be used. Wells that are inoculated too lightly may give false-negative reactions.

MATERIALS REQUIRED BUT NOT PROVIDED

Standard microbiological supplies and equipment such as loops, swabs, applicator sticks, other culture media, incinerators, and incubators, etc., as well as serological and biochemical reagents, are not provided.

QUALITY CONTROL

Test Organisms Well Incubation Results
Time Temperature Atmosphere
Candida glabrata
ATCC ® 66032
Control

Trehalose


Maltose


Sucrose







2hr 35°C Aerobic Blue/blue-green

Positive,
yellow/greenish-yellow

Negative,
blue/blue-green/green

Negative,
blue/blue-green/green








Saccharomyces cerevisiae
ATCC ® 9763
Control

Trehalose


Maltose


Sucrose







2hr 35°C Aerobic Blue/blue-green

Negative,
blue/blue-green

Negative,
blue/blue-green

Positive,
yellow/greenish-yellow








USER QUALITY CONTROL

Physical Appearance

GlabrataQuick™ Kit

GlabrataQuick™ Kit (Cat. no. Z298). Candida glabrata wil be positive for trehalose and negative for both the maltose and sucrose wells.

GlabrataQuick™ Kit

Showing Non- C. glabrata results for the GlabrataQuick™ Kit (Cat. no. Z298). If the maltose and/or sucrose well(s) is positive then the isolate is not Candida glabrata , regardless of the trehalose reaction.


GlabrataQuick™ Kit

Showing all results for the GlabrataQuick™ Kit (Cat. no. Z298).


RefeRENCES

1. Jorgensen., et al. Manual of Clinical Microbiology, American Society for Microbiology, Washington, D.C.

2. Tille, P., et al. Bailey and Scott's Diagnostic Microbiology, C.V. Mosby Company, St. Louis, MO.

3. Isenberg, H.D. Clinical Microbiology Procedures Handbook, Vol. I, II & III. American Society for Microbiology, Washington, D.C.

4. Koneman, E.W., et al. Color Atlas and Textbook of Diagnostic Microbiology, J.B. Lippincott Company, Philadelphia, PA.

5. Quality Assurance for Commercially Prepared Microbiological Culture Media, M22. Clinical and Laboratory Standards Institute (CLSI - formerly NCCLS), Wayne, PA.

6. Stockman, L. and G. Roberts. 1985. Abstracts of Annual Meeting of American Society of Microbiology, p.377 (F-80). Washington, D.C.

7. Frye, K.R., J.M. Donovan, and G.W. Drach. 1988. Torulopsis glabrata urinary infections: a review. J. Urol.; 139:1245-1249.

8. T. Scognamiglio, Lawrence P. and D.H. Larone. 2004. Evaluation of a New commercially Available Rapid Assimilation of Trehalose (RAT) Test for the identification of Candida glabrata. Abstracts of the General Meeting of the American Society for Microbiology, New Orleans, LA.

9. Joann, P.F., et al. 1999. Comparision of Four Methodologies for Rapid and Cost Effective Identification of Candida glabrata. Journal of Clinical Microbiology.

10 A.M. Freydiere., et al. 2003. Rapid Identification of Candida glabrata with a New Commercial Test, GLABRATA RTT. Journal of Clinical Microbiology.

11. Michael, BS., et al. 1999. Comparative Performance of the RapID Yeast Plus System and API 20C AUX Clinical Yeast System. Journal of Clinical Microbiology.


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