Gamma Glutamyl Transferase Reagent (5X Concentrate)
Gamma Glutamyl Transferase Reagent (5X Concentrate)
Gamma Glutamyl Transferase Reagent (5X Concentrate)
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<strong>Gamma</strong> <strong>Glutamyl</strong> <strong>Transferase</strong> <strong>Reagent</strong><br />
(<strong>5X</strong> <strong>Concentrate</strong>)<br />
V<br />
PRODUCT SUMMARY<br />
Stability : 14 days at 2-8°C<br />
Linear Range : Up to 600 U/L<br />
Specimen Type : Serum<br />
Method : Kinetic<br />
<strong>Reagent</strong> Preparation : Add 7 mL of distilled or<br />
deionised water.<br />
INTENDED USE<br />
This reagent is intended for the in vitro quantitative determination gammaglutamyltransferase<br />
(GGT) [(γ-<strong>Glutamyl</strong>) - Peptide: Amino Acid γ-<strong>Glutamyl</strong>transferase,<br />
EC2.3.2.2], in human serum.<br />
SUMMARY<br />
Although gamma-glutamyltransferase (GGT) is present in a variety of tissues, the<br />
serum enzyme appears to be primarily from the hepato-biliary system. Consequently,<br />
GGT is elevated in all forms of liver disease or damage. It is clinically useful in<br />
detecting obstructive jaundice, cholangitis, and cholecystitis.<br />
Elevated levels are also observed with drug use (alcohol, sedatives, anticonvulsants<br />
and tranquilizers). 1<br />
PRINCIPLE<br />
The first commercially available kinetic methods for the determination of GGT were<br />
based on the work of Szasz 2 , Rosalki and Tarlow 3 . These methods utilised γ-glutamylp-nitroanilide<br />
(Glu-4-NA) as the substrate, however the poor solubility and stability of<br />
Glu-4-NA was a major limitation. In order to improve the method Persijn 4 investigated<br />
further with Glu-4-NA derivatives and found that γ-glutamyl-3-carboxy-4-nitroanilide<br />
(Glucana) was superior to the Glu-4-NA with respect to both solubility and stability.<br />
The Glucana substrate now forms the basis of the IFCC and ECCLS recommended<br />
procedures. The GGT- Soluble Substrate method utilises Glucana in a one step<br />
method in which the reaction is initiated with the addition of sample. GGT present<br />
in the sample catalyzes the transfer of the glutamyl group from the substrate to<br />
glycylglycine forming glutamylglycylglycine and 5-amino-2-nitrobenzoate.<br />
L-γ-glutamyl-3-carboxy-4-nitroanilide + glycylglycine<br />
<br />
SYMBOLS IN PRODUCT LABELLING<br />
For in vitro diagnostic use<br />
Batch code/Lot number<br />
Catalogue number<br />
Consult instructions for use<br />
Xn - Harmful<br />
STORAGE AND STABILITY<br />
1. The unopened reagents are stable until the expiration date stated on the label<br />
when stored at 2-8°C.<br />
2. After reconstitution the reagent is stable for 14 days when stored on the Olympus<br />
Demand or Reply at 2-8°C.<br />
DETERIORATION<br />
1. The dry powder reagent should be a white to off-white, free flowing powder.<br />
2. The reconstituted reagent should be a clear, slightly yellow solution.<br />
3. Failure to achieve assay values on freshly prepared control sera could indicate<br />
deterioration.<br />
SPECIMEN COLLECTION<br />
Clear, non-hemolyzed serum is the recommended sample. 5<br />
SAMPLE STORAGE<br />
GGT is stable for 7 days when stored at 2-8°C. 6<br />
INTERFERING SUBSTANCES<br />
1. No interference from Lipemia, measured as triglycerides, up to 200 mg/dL.<br />
2. No interference from Conjugated Bilirubin up to 60 mg/dL.<br />
3. No interference from Free Bilirubin up to 48 mg/dL.<br />
4. No interference from Hemoglobin up to 350 mg/dL.<br />
5. Young has reviewed drug effects on serum GGT levels. 6<br />
PROCEDURE<br />
Test Parameters<br />
Refer to the Thermo <strong>Reagent</strong> Applications.<br />
Temperature Limitation<br />
Use by/Expiration Date<br />
CAUTION. CONSULT INSTRUCTIONS<br />
FOR USE.<br />
Manufactured by<br />
GGT<br />
MATERIALS PROVIDED<br />
GGT <strong>Reagent</strong><br />
10 x 7 mL<br />
L-γ-glutamylglycylglycine + 5-amino-2-nitrobenzoate<br />
The rate of formation of 5-amino-2-nitrobenzoate is proportional to the activity of GGT<br />
present in the sample and can be measured bichromatically at 410/660nm.<br />
REAGENTS<br />
Reactive Ingredients Initial Conc. Final Conc.<br />
Tris buffer 550 mmol/L 110 mmol/L<br />
Glycylglycine 550 mmol/L 110 mmol/L<br />
L-γ-glutamyl-3-carboxy-4-nitroanilide 16 mmol/L 3.2 mmol/L<br />
Also contains non-reactive fillers and stabilizers<br />
pH 8.20 ± 0.1 at 20°C<br />
PRECAUTIONS<br />
Do not ingest. Avoid contact with skin and eyes. If spilt thoroughly wash affected<br />
area with water. <strong>Reagent</strong> contains sodium azide which may react with copper or<br />
lead plumbing. Flush with plenty of water when disposing. For further information<br />
consult the Bicarbonate <strong>Reagent</strong> Material Safety Data Sheet. The Packaging of<br />
This Product Contains Dry Natural Rubber.<br />
R22 Harmful if swallowed.<br />
R36/38 Irritating to eyes and skin.<br />
S26 In case of contact with eyes, rinse immediately with plenty of water<br />
and seek medical advice.<br />
REAGENT PREPARATION<br />
Add 7 mL of freshly distilled or deionized water to each vial. Swirl gently to dissolve<br />
the contents. Allow to stand 10 minutes at room temperature before use.<br />
MATERIALS REQUIRED BUT NOT PROVIDED<br />
1. Clinical chemistry analyser capable of maintaining constant temperature<br />
(37°C), using 5x concentrated reagents and measuring absorbance at<br />
405 nm (405 - 420 nm)<br />
2. Thermo <strong>Reagent</strong> Applications for the Olympus Demand or Reply.<br />
3. Thermo Data-Trol N and Data-Trol A (Cat. No. 1902-050 or TR40001 and<br />
1901-050 or TR41001) or equivalent.<br />
STABILITY OF FINAL REACTION MIXTURE<br />
The instrument automatically computes every determination at the same time<br />
interval.<br />
CALIBRATION<br />
A calibrator is not required for the GGT Procedure. One U/L of GGT activity is that<br />
amount of enzyme which produces one mmol/L of p-nitroaniline per minute.<br />
LINEARITY<br />
Linearity extends to 600 U/L. Samples exceeding linearity should be diluted with<br />
normal saline and repeated. Multiply the result by the dilution factor when calculating<br />
the unknown.<br />
QUALITY CONTROL<br />
Normal and abnormal control sera of known concentrations of GGT should be<br />
analyzed routinely with each group of unknown samples. Data-Trol N and Data-Trol<br />
A (Cat. No. 1902-050 or TR40001 and 1901-050 or TR41001) are recommended<br />
for this purpose.<br />
CALCULATION OF RESULTS<br />
Results, expressed as U/L at 37°C, are automatically calculated.
LIMITATIONS<br />
See Storage and Stability, Deterioration, Specimen Collection, Interfering Substances,<br />
Sample Storage, and Linearity sections for limitations to this procedure.<br />
EXPECTED VALUES 3<br />
At 37°C : Males: < 50 U/L<br />
Females: < 30 U/L<br />
The quoted values are representative of the expected range for this method and<br />
should serve as a guide only. It is recommended that each laboratory verify this<br />
range or derives a reference interval for the population that it serves. 7<br />
PERFORMANCE CHARACTERISTICS<br />
Precision<br />
WITHIN-RUN Level 1 Level 2<br />
No. of data points 20 20<br />
Mean U/L 20 67<br />
SD 0.41 1.00<br />
CV% 2.05 1.49<br />
BETWEEN DAY Level 1 Level 2<br />
No. of data points 20 20<br />
Mean U/L 22 67<br />
SD 1.80 2.05<br />
CV% 8.18 3.06<br />
COMPARISON STUDIES<br />
A comparison of the Thermo GGT reagent (y) with a commercial reagent of the same<br />
methodology (x) was performed on 147 human samples in a range of 2 - 831 U/L.<br />
A correlation coefficient of 0.999 was obtained; the linear regression equation was<br />
y = 0.99 x + 2.7.<br />
ANALYTICAL SENSITIVITY<br />
Based on an instrument resolution of A= 0.001, this Thermo GGT Procedure has<br />
a sensitivity of 2.6 U/L.<br />
REFERENCES<br />
1. Kachmar JF, Moss DV. “Enzymes” in Fundamentals of Clinical Chemistry. Tietz<br />
NW (Ed) WB Saunders Co. Philadelphia 1976; page 621-3.<br />
2. Szasz G. Clin Chem 1969; 15: 124-36.<br />
3. Rosalki SB, Tarlow D. Clin Chem 1974; 20: 1121-4.<br />
4. Persijn JP and van der Slik W. J.Clin.Chem.Clin Biochem. 1976; 14: 421-7.<br />
5. Tietz, N.W., Clinical Guide to Laboratory Tests, W.B. Saunders, Philadelphia,<br />
1983, p. 244.<br />
6. Young DS. Effects of Drugs on Clinical Laboratory Tests. Third Edition. 1990;<br />
3: 183-5.<br />
7. Wachtel M et al, Creation and Verification of Reference Intervals. Laboratory<br />
Medicine 1995; 26:593-7.<br />
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Phone: (03) 9790 4100<br />
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U.S.A.<br />
Phone: (800) 558 9115<br />
Fax: (412) 788 6833<br />
Reorder Information and Technical Support<br />
Catalogue No.<br />
Configuration<br />
7200-016A 10 x 7 mL<br />
Australia International U.S.A.<br />
Phone 1800 333 110 61 3 9790 4100 (800) 558 9115<br />
Facsimile (03) 9790 4155 61 3 9790 4155 (412) 788 6833<br />
P/N: PI191041.01 Rev 12/2006