Biochemistry

I '

y GLUTAMYL TRANSFERASE KIT /&-{ (Carboxy Substrate Method) M 1:{ IJ..~ -

CE (For invitro diagnostic use only)

Appropriate biosafety practices should used for materials that INTENDED USE . . .~er-are suspected of containing infectiOUS agents. . yGlutamyiTransferaseKitisusedforthedetenn1nabonofy-Glutam,.-----.H•~ di... . solid and liquid waste and test components m

, A"': ·1y. an e spoomens, 'd 1• s M29 or Trans,erase .,uvI In serum. accordance with local regulations and NCCLS gu1 e 1ne ,

SUMMARY y Glutamyt Transferase (GGT) is an enzyme found mainly in serum from hepatic origin, though the highest levels are. in k!dneys. Elevat~d levels are found in hepatobiliary and pancreatic diseases, chronic alcoholism, myocardial infarction with secondary liver damage and diabetics.

PRINCIPLE GGT catalyzes the transfer of amino group between L-y-Glutamyl-3- carboxy-4-nitroanilide and Glycylglycine to form L-y- , Glutamylglycytglycine and 5-amino-~-nitrobenzoate. ~e rate ?f fonnation of 5-amino-2- nitrobenzoate Is measured as an increase In absorbance which is proportional to the GGT activity in the sample.

,::r~. _. GGT . L-y-Glutam~-~rboxy4-nitroanilide-+L-y-Glutamylglycytglycine

...... -·. + - Glycylglycine

EXPECTED VALUES Serum (Males)

(Females)

. + 5-amino-2-nitrobenzoate

: 10-50 U/Lat37'C : 7-35 U/Lat37'C

It is recommEJ11ded that each laboratory establish its own nonnal range representing its patient population.

--- PRESENTATION- · ___ , - -- - --- -- ·-·· --- . - .

!REF! Pack Size

L 1 Buffer Reagent T1 Substrate Tablets

COMPOSmON

1102100102 10 x 2 ml

25ml 10Nos.

Tris Buffer 110 mM; pH 8.5; Glycylglycine 110 mM; Carboxy Substrate_ (Glupa) 4 mM.

STORAGE/ STABILITY Contents are stable at 2-8'C till the expiry mentioned on the labels.

REAGENT PREPARATION Working reagent: Dissolve 1 Substrate Tablet in 2.2 ml of Buffer Reagent This wooong reagent is stable for at least 15 days when storedat2-8'C.

SAMPLE MATERIAL Serum. Free from hemolysis. GGT is reported to be stable in serum for 3daysat2-8°C.

SAMPLE WASTE AND DISPOSAL Do not reuse the reagent containers, bottles, caps or plugs due to the risks of contamination and the potential to compromise reagent performance. . . This product requires the handling of human specimens. It Is recommended that all human sourced material are considered potentiany hazardous and are handled in accordance with the OSHAstandard on blood borne pathogens.

other published biohazard safety guidelines.

MATERIALS REQUIRED BUT NOT PROVIDED . Photometer analyzer with standard the~ostatic cuvette holder, micropipette and appropriate laboratory equipment. •

PROCEDURE Wavelength I filter Temperature Light path

: 405nm 37'C/30' C/25"C

: 1 cm

Pipette into a dean dry test tube labelled as Test (T):

Addition Sequence (T)(ml)

Working Reagent 1.0

Incubate at the assay temperature for 1 minute and add

Sample 0.1

Mix well and read the initial absorbance A, after 30 secs. and repeat the absorbance reading after every 1 & 2 mins. Calculate the mean absorbance change per min. ( 6A / min).

CALCULATIONS GGT Activity in U/L = 6A /min.x 1158

QUAUTYCONTROL - - - ------ The following process is recommended for QC during the assay of GGT. •Define and establish acceptable range for your laboratory. 1. Two levels of control.(Nonnal and Abnonnal) are to be run on a

daily basis. , 2. If QC results fall outside acceptance criteria, recalibration may be

necessary. 1'

3. Review QC results and run acceptance criteria following a change of reagent lot.

SPECIFIC PERFORMANCE CHARACTERISTICS Linearity: The procedure is linear upto 700 U/L at 37' C. If the absorbance change (Ml min) exceeds 0.250, use only the value of the first 2 mins. to calculate the result, or dilute the sample 1 + 9 with nonnal saline (NaCl 0.9%) and repeat the assay (Results x 10).

Limit of detec1ion: The limit of detection for GGT is 1 U/L.

Interferences: For diagnostic purposes, the results should always be assessed in conjunction with the patient's medical history, clinical examination and other findings.

<~

30~,q<- - .

Acla1 t1on Sequence

REFERENCES · , . 1. IFCC methods for the measurement of catalytic concentrations of precision: . precision studies were performed with two ~ntrols using NCCLS protocol EP5-A. The results of the precision studies are shown below:

enzvmes,J.Clin. Chem. Clin Biochem,(1986)24:4~7 . . 2. Clinical Chemistry, Principles procedures Correlations, Michael L

Sample Within-run Between-run Total

Mean CV% Mean CV'/o Mean CV%

--37.38 - 5a18- --39.87- -- --

Control1 ·'" v,a,

1.52 231.4 1.05 436.8 2.57

Bishop, et. al., 5th Edition. 3. Data on file: Coral Clinical Systems.

Reaction : Kinetic Interval : 30 Sec. : 0.10ml

System Parameters

ControI2 205,4 Wavelength : 405 nm Sample Vol. Zero Setting : Distilled Water Reagent Vol. : 1.00 ml

Method comparison: lncub. Ttmp. : 37° C Standard - : 1158 Comparative studies were done to compare our reagent with another

commercial y Glutamyl Transferase Assay. No significant differences were observed. Details of the comparative studies are available on

lncub. nm• DelaY,Tlmi·

: - Factor : 30 Sec.

Reacl Slope : Increasing : 700 U/L

request

TEMPERATURE CONVERSION FACTORS

Assay Temperature Desired Reporting Temperature 25'C 30'C 37'C

25'C 1.00 1.31-- 1.79

30'C 0,73 1.00 1.30

37'C 0,56 0.77 1.00

NOTE ·•· - , -- ' ---invifrooiagnosficreage'nfforlaboratory and profession~! use only Not - -

for medicinal use. The reagent contain sodium atide 0.1 % as preservative. Avoid contact with skin and mucosa. pn djsposal flush with large quantities of water. Only clean and dry giass~e nj.Jst be used. Samples having a very high activity show a very high initial absorbance. If this is suspected then dilute the sample and repeat the assay. The reagent may be used in several automated analyzers. Instructions are available .o.n rpquest. Do not use turbid, oeteriorated or leaking reagenis. ,/ ,, ., ..

Read Time : 120 Sec. No. otriad. : 4

%re . • llVD! In vltTD

%'C Store at 2-S'C ·Manufacturer Diagnostic Medical Device

Uaeby []l1 (ustday af a1al8d month)

E] Date ol !REF! Manufacture

Consult I LOT I Batch Number lnstiuctkins for use

C&talogue I EC I REP I Number Aut,at\Nd Repmentallve

In tho EIIOl)Ul1 Comnu,ity

...a MaM!factured by:

Coral Clinical Systems A Division of Tulip Diagnostics (P) Ltd.

Unearlty Units

@] Buffer Reage11t

Substrate Tablets'

' . Pl.OT NO. M-46, PHASE Ill B, VERNA IND. EST., VERNA, GOA-403 722, INOIA. .

REGD .. OFFICE: GITANJALI, TULIP BLOCK. DR. ANTONIO.DO REGO BAG~- '' .AL TO SANTA~RUZ, BAMBOLIM COMPLEX P.O., GOA-403 202, INDIA. ,- :- . .

I EC I REP I CMC Medical Devices & Drugs S.L.,

Ci Horacio Lengo No. 18, CP 29006, Malaga, Spain.

wz • se. a •*Ml ooW&fiiU!::.a.aweWiW

: U/L

I Carboxy Subslratt I Carboq~

Method

tl Thlswrjup

b~c,t.;a, r& ,1hn1 lfl ,1b$0rbanLe

'YrllFS J Cl,r

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