LIQUID LINE -G-test - Wiener lab.

1 LIQUID line . -G-test cin tica AA. C Modified Szasz method for the determination of - glutamyl transferase in serum or plasma. Substrate recommended by the IFCC. SUMMARY Monoreagent (premixed): stable for 4 weeks at 2-10oC. - glutamyl transferase ( -GT) is a membrane enzyme widely Protect from sunlight. distributed in the body. It is primarily located in the kidney, seminal vesicles, pancreas, liver, spleen and brain. Its activity INSTABILITY OR DETERIORATION OF REAGENTS. is influenced by any factor altering the cellular membranes of When the spectrophotometer has been set to zero with dis- the organs that contain it. In the case of liver disorders, -GT tilled water, absorbance readings of the Monoreagent higher generally indicates toxic aggression. However, its determina- than indicate reagent deterioration. tion only has clinical relevance when its values are compared to those of other greater organ-specificity enzymes. SAMPLE. -GT determination together with alkaline phosphatase, Serum or plasma transaminase and bilirubin, significantly broadens the spec- a) Collection: obtain sample in the usual way.

2 Trum for differential diagnosis of primary and secondary liver b) Additives: when using plasma, collect it with EDTA as diseases, being part of the hepatic profile. anticoagulant. c) Known interfering substances: no interferences have been PRINCIPLE observed by bilirubin up to 280 mg/l (28 mg/dl), triglycerides up - glutamyl transferase is a carboxypeptidase that catalyzes to g/l (540 mg/dl) nor hemoglobin up to g/dl (390 mg/dl). the following reaction: See Young, in References for effect of drugs on the -GT present method. L- - glutamyl -3-carboxy-4-nitroanilide + glycilglycine d) Stability and storage instructions: -GT in serum is L- - glutamyl glycilglycine + 5-amino-2-nitrobenzoate stable for 2 weeks at 2-10oC and up to 6 months at -20oC. PROVIDED REAGENTS REQUIRED MATERIAL (non-provided). A. Reagent A: Tris buffer solution containing glycilglycine. - Spectrophotometer. B. Reagent B: L- - glutamyl -3-carboxy-4-nitroanilide solution. - Micropipettes and pipettes for measuring the stated vol- umes.

3 Final concentrations - Spectrophotometric square cuvettes. Tris 100 mmol/l; pH - Water bath at selected assay temperature. L- > mmol/l - Stopwatch. 100 mmol/l ASSAY CONDITIONS. NON-PROVIDED REAGENTS - Wavelength: 405 nm Saline solution (9 g/l NaCl). - Reaction temperature: 25, 30 or 37oC. - Reaction time: 3 minutes INSTRUCTIONS FOR USE - Sample volume: 100 ul Provided Reagents: ready to use. They may be used sepa- - Reagent B volume (Substrate): 1 ml rately or as Monoreagent, mixing 4 parts Reagent A with - Final reaction volume: ml 1 part Reagent B ( 4 ml Reagent A + 1 ml Reagent B). WARNINGS PROCEDURE. Reagents are for "in vitro" diagnostic use. In a cuvette kept at the selected temperature, place: Use the reagents according to the working procedures for clinical laboratories. Monoreagent 1 ml The reagents and samples should be discarded according Pre-incubate a few minutes. Then add: to the local regulations in force. Sample 100 ul STABILITY AND STORAGE INSTRUCTIONS Mix at once and continue incubation immediately and Provided Reagents: are stable at 2-10oC until the expiration simultaneously start stopwatch.

4 Record absorbance at 1, date stated on the box. 867028570 / 03 p. 1/3. 100 ml: - 4 x 20 ml Reagent A. 2 and 3 minutes. Determine average absorbance change - 1 x 20 ml Reagent B. ( A/min) subtracting each reading from the previous one (C d. 1009330). and averaging values. Use this mean for calculations. 100 ml: - 4 x 20 ml Reagent A. - 1 x 20 ml Reagent B. CALCULATIONS (C d. 1009258). - glutamyl Transferase (U/l) = A/min x 1,158 125 ml: - 5 x 20 ml Reagent A. - 2 x 12,5 ml Reagent B. SI SYSTEM UNITS CONVERSION (C d. 1009616). -GT (U/l) x = -GT (ukat/l). REFERENCES. QUALITY CONTROL METHOD - Lum, G.; Gambino, - Clin. Chem. 18:358 (1972). Each time the test is running, analyze two levels of a qual- - Burrows, S.; Feldman, W.; McBride, F. - Am. J. Clin. Path. ity control material (Standatrol S-E 2 niveles) with known 64/3:311 (1975). - glutamyl transferase activity. - Szasz, G. - Clin. Chem. 15/2:124 (1969). - - Scand. J. Clin. Lab. Invest. 36:119 (1976).

5 REFERENCE VALUES - Szasz G.; Weinmann G.; Staler F.; Wahlefeld W.; Persijn Temperature 25oC 30oC(*) 37oC(*) J. - Z. Klin. Chem. Biochem. 12:229 (1974). Men 6-28 U/l 8-38 U/l 11-50 U/l - - J. Clin Chem. Clin. Biochem. 21: 633 (1983). Women 4-18 U/l 5-25 U/l 7-32 U/l - Young, - "Effects of Drugs on Clinical Laboratory Tests", (*). Calculated AACC Press, 5th ed., 2000. - CLSI: Clinical and Laboratory Standards Institute (ex- PROCEDURE LIMITATIONS NCCLS) - Protocols EP-15A, 2001 / EP-17A, 2004. See Known Interfering Substances under SAMPLE. PERFORMANCE. a) Reproducibility: CLSI protocol EP15-A was applied. Two activity levels were tested, in replicates by four, during 5 days. With the obtained data, total and intra-assay preci- sion were calculated. Intra-assay precision (n = 20). Level U/l U/l %. U/l U/l %. Total precision (n = 20). Level U/l U/l %. U/l U/l %. b) Linearity: manually, the reaction is linear up to A/min (250 U/l). For higher values, dilute sample 1/5 or 1/10 with saline solution and repeat assay under the same assay conditions.

6 Multiply results by the dilution performed. In autoanalyzers, a linearity up to 1200 U/l may be observed. c) Analytical sensitivity: depends on the photometer used and wavelength. In spectrophotometer with 1 cm optical length square cuvettes, for A/min of , the minimum detectable activity change will be of 1 U/l. PARAMETERS FOR AUTOANALYZERS. For programming instructions check the user manual of the autoanalyzer in use. Wiener LAB. PROVIDES. 100 ml: - 4 x 20 ml Reagent A. - 1 x 20 ml Reagent B. (Cat. 1421404). 867028570 / 03 p. 2/3. Symbols The following symbols are used in packaging for Wiener lab. diagnostic reagent kits. C. This product fulfills the requirements of the Euro- pean Directive 98/79 EC for "in vitro" diagnostic M Manufactured by: medical devices P. Xn Authorized representative in the European Com- munity Harmful V "In vitro" diagnostic medical device Corrosive / Caustic X Conteins sufficient for <n> tests Xi Irritant H Use by i Consult instructions for use l Temperature limitation (store at).

7 Do not freeze Calibr. Calibrator b Control F Biological risks Volume after reconstitution b Positive Control Cont. Contents c Negative Control g Batch code h Catalog number M Wiener Laboratorios Riobamba 2944. Wiener lab. 2000 - Rosario - Argentina Dir. T c.: Viviana E. C tola Bioqu mica Registered product PM-1102-19 2000 Rosario - Argentina 867028570 / 03 p. 3/3 UR120831.