Erba’ €rbaLisa® Total Triiodothyronine (tT3) 96 Tests Product Code: 133321 Erbalisa® TOTAL TRIODOTHYRONINE (tT3) ELISA KIT IS INTENDED TO BE USED FOR THE QUANTITATIVE MEASUREMENT OF TOTAL ‘TRIIODOTHYRONINE (¢T3) IN HUMAN SERUM. INTRODUCTION ‘otal Triadothyronine (73) is 2 useful marker for the diagnosis of hypothyroidism and hyperthyroidism. The level of t73 is decreased in hypothyroid patients and is increased in hyperthyroid patients, Graves disease and pregnancy, THYROID HORMONES PRINCIPLE Erbalisa® Total Tiiodothyronine [{T3} is 9 sold phase competitive EUSA, The samples, assay buffer and T3 enzyme conjugate are added to the wells coated with ant-T3 monoclonal antibody. T3 inthe patents serum competes with the T3 enzyme conjugate for binding sites. Unbound 73. and 13 enzyme conjugate is washed off by washing buffer Upon the additon ofthe substrate, the intensity of color is inversely proportional to the. urve i plotted relating coor intensity to the concentration of the T3, concentration of 1T3in the samples. Using 6 standard kar conrenrs z [see Reogens ter ae Ceatedwirovels-idlaway mires & 73 StndoasReodytowe fas | 6.05 | conugte 13 ennme compat oncnioe, in [bbe recosthed) [sah Dilvent=T3 assay divert oe : 1 vale 2 ml ~ color Reagent - TMi substrate ready to ae : 1 botle 2 Stop sohton = Resdy ose ae 1 bote2 ml Wash oncertate (20s) = ite 120 ith ed water . [tote 25 | Black od exposure to ight during incubation [eprrned “[ Pte Seater To cover tis during incubation, to reverts due o apoaion ae Zip Lock Bag To store unused microwells a ‘No. NOTE: tems 1 to 7 should be stored at 28°C. MATERIALS REQUIRED BUT NOT PROVIDED 1. Disposable gloves and protective glasses, 2. Distiled or deionized water 3, Precision pipe 4. Disposable pipette tips 5. Precision EUSA reader capable of reading absorbance at 450.nm (Etbs 6 2 8 Automated microplate washing instrument (Erba LisaWash I Measuring cylinder Absorbance paper or paper towel == \ kei STORAGE AND SraBiLiTy Keep the unused meronell sealed in a dry bag with desccats gual TRANSASIA BIO-MEDICALS LTD.3 Erba’ ‘The reagents are stable unt expiration ofthe kt Donor expose test reagents 1 eat, sun a tong ight WARNINGS AND PRECAUTIONS ental bahnadovs mate: The standards contin human ouce components ed at dane fees sone els antibody with FOA licensed reagents. However there are no hod ; f somes oal tal teak 8 ther inf agent: absent, These reagents should be handled at the Biosafety Level 2, a: ‘exommendedin the Centers for Doras ConroVNatonl Insttes feat manval“Bosultyinhcrbioemeal and medial cherie a it its are handled, tpt by moth. Danske ea, or nk in the areas n which specimens ot eagents THeconporoe tna manele scaton ne eae ohare, Iisreonmended that standards and serum samples Deon dane eet ‘oma resus wil be obiined by tet adherence to ths protoclAccUrte and rece pipetting, 3 wells following tengeraurerequremens prscred ae exert AnyGenston toms hav veld Gon Drab gloves should be worn throughout the proces ‘saya be performed by rained personne ony. forinvivodeenomt use ony rar to dota allwaste mater should be collected and kent in Sx sodium hypochlorite cation or 30 minutes Donat use thet bron its beled expry ate Doro interchange reagents been een se clear serum. arate mate sha be removed by centration Use separate tis or stands and india et pecmene, 0 not expose coor eagen to sunignt Dates oF deo wat" tobe urd for lution of washing bute touching TNE Maro es ele slong wh atts seb et in ele conn ah phone 2ipock bag, ‘SAMPLE COLLECTION AND HANDLING 1. 1 Collect blood specimens and separate the serum immediately, 5 Ree may be stored reigerates at (28 das storage tne exceeds 5 day, tae foven 3 (-20°C) for up to ane month, 3. Avoid mutiple freeze-thaw eyces {Brito assay, frozen sera shouldbe completely thawed and maxed well 5. Donot use gros lipemic specimens, : —_ 1. TB-enzyme Conjugate Solution Tae T3-eteyme conjugate 1:1 with conjugate dentin suitable contain ‘TSteatet should be used within wentyour hous for macanom serie Wels) a nce ofthe assy. I required, store at 2-8. a2 | «ae ry a [ere 360u! | 400 ul | aa0 at | a80ur 24m | 28m | 32m | 36m 7 Conjugate | 0.4 mi 40m | 4m | 48m sr | | Omi | 44m) | 48m 2 Wash Buter Pepare wash buflerby adding the contents ofthe bot (25 m, 20010475 mio ited or donee mater Stor at oom temperature (20.25°¢) ‘ASSAY PROCEDURE 2 eight sansa sos essen 4.66 100 yl of T3-en2yme Conjugate solution to all wells (see i section) 0a wes Regent reparation 5. Swirl the microplate gently for. 20-30 seconds to mix and cover, teil : 6. Inte Somes stoom coe 2. Remove a om a wee Wash al etn @ Addionyartie subsvate soliton toate Wes with 300 u! see wash butfer Preparation). Blot ‘on absorbent paper towels. 9. _Incibtestoom enperuretor is oa 10. ‘AidSoult top sclton cach wat ny, hl and ety mio 15.20 sends 2S Rebs sinncn Sheer Mints afer ad the top oluton cALcuLAnON oF resus The standard curves tobe constructed as follows 1 2 hack T3 standard ale on each standard vl This vay or ofthe standart atacnce “IBM vary fom lot tlt. Make sure you check the value on every kt. See example tua] TRANSASIA BIO-MEDICALS Lrp, 5-11, OIDC, Ringanwade, Daman ~ 396 210, india.graph paper Draw the best curve through the points 3, Read the absorbance for cantrals and each unknown sample [EXAMPLE OF A STANDARD CURVE PERFORMANCE CHARACTERISTICS 1. Sensitivity ‘The Sensitity of 13 ELISA was assessed by cunning 24 repicat response curve Serum 2. Precision 173 Cone. (ng/ml) tes of “0 Standard inthe assay and re-poting the #250 from the mean of of o trom Curve. Record the Standard 00a 450.nm | 2587 0 | 1974 os 1282 15 0.730 3 0.340 6 | 0.185 J the dose [Mean + 250 (Sensitivity) _| ‘Standard 0.066 ng/ml | of each of thee sera; normal, low and heh. 1. Intra-Assay Precision was determined by assaying 16 replicates [__ Cafe o variation ~ Serum | No. of Repliates ‘ean mg/m Epic eer ete 25 69% era ee fice eeciae ce Peree bal Pore 3 1 16 442 0.13 3.2% af aicsserum polsn 16 separate ars, singa standard cave consuced fo each bb inter assay Precision was determined by assaying duphicat “ean nant [Standard Deviation Coefficient of Variation (4) 0109 "asi [os fem 4 aus8 7% 3. Specificity “The cross reaction of concentrations, The the same amount of tracer oss eactwity was calculated by derivinga 2 analyte Po — (eee Seats <—cos ce EXPECTED VALUES iestudy of euthyroid adult population was vrcnanon and expected ranges of samples ae presente inthe follow (105 samples) 1186 (0334 5 ng/ml | Mean standard deviation. ual the Thyroxine antibody to selected substances wa Tes concentaon Tested Undertaken to determine expected values for the total T ing substance to a serum mati a various 1 evaluated by adding the interfer ee neen dose of interfering substance to dose of Thyroxine needed to displace 3 £18 test system, The mean value, standard 05): ing table (total samples test’ TRANSASIA BIO-MEDICALS LTD. B-11, OIDC, Ringanwada, Daman - 396 210, India,| _ Erba? \TIONS OF THE TEST : ie Its obtained using this kit serve only as an aid to diagnosis and should be interpreted in relation to the patient's history, physical 1. The test results obtained using : findings and other diagnostic procedures 2. Do not use sodium azide as preservative. Sodium azide inhibits HRP enzyme activities. LIMITED EXPRESSED WARRANTY DISCLAIMER Transasia Bio-Medicals Ltd. products are warranted to meet the applicable product specifications described. Notice of non-conforming products should be made to Transasia Bio-Medicals Ltd for which lability is limited to either replacement of the product or refund of the purchase price of the product and in no case liabe to for claim of any kind for an amount greater than the purchase price of the goods in respect of which damages are likely to be claimed. Transasia Bio-Medicals Ltd, disclaims any and all responsibility for any injury or damage or legal implications which may be caused by the fault of the user or the buyer in accordance with the limitations and specifications here in. Due to continuous development, the manufacturer reserves the right to improve/change any specifications/components without prior information/notice to the buyer. REFERENCES: 1. Agheranya JC. Clinical usefulness of ELISA technique in the assessment of thyroid function. West Afr J Med 1990; 9(4):258-63. 2. Baumgartner Parzer SM; Wagner L; Reining G; Sexl V; Nowotny P; Muller M; Brunner M; Waldh“ausl W. Increase by tri-iodothyronine of endothelin-1, fibronectin and von Willebrand factor in cultured endothelial cells. J Endocrinol 1997; 154(2):231-9, 3. Maes M; Mommen k; Hendricks D; Peeters D; O’Hondt P; Ranjan R; De Meyer F; Scharp’e S. Components of biological variation, including Seasonality, in blood concentrations of TSH, tT3, 4, PRL, cortisol and testosterone in healthy volunteers, Clin Endocrine (Oxf) 1997; 46(5):587- 98, 4. Santini; ChiovatoL; Bartalena ; Lapi; Palla R; PanichiV; Velluzzi ; Grasso L; Chopra; Martino €; Pinchera A Study of serum 3,5,3’-triiodothyronine sulfate concentration in patients with systemic non-thyroidal illness. Eur J Endocrinol 1996;134(1):45-9,8 €rbaLisa® Total Triiodothyronine (tT3) Assay Procedure Product Code: 133321 Define Standards/Sample Placement Label the microwells for T3 standards and samples, in Addition ia duplicate Standards/Samples Addition eee Add 50 ul of standards and samples in the wells, as marked. = Lf lal , Add 100 ul of freshly prepared T3 Conjugate Solution . / c T3 enzyme conjugate solution to all the wells, = = . a Swirl the microwells gently for Shaking 20-30 seconds to mix, and cover = a with plate sealers, Incubate the microwells for 60 Incubation minutes at room temperature = (20-25°c) : 300 pl/well Washing me teg |e 3 cycles; Blot on the tissue paper at the end Substrate/Color Reagent 4 7 7 ‘Add 100 ul TMB substrate (RTU) Addition to each well Incubate the microwells for 15 Incubation minutes at room temperature (20-25°¢). Stop solution Addition Add 50 yl of stop solution (RTU) to each well, Reading OD Read the OD of microwells at 450 nm within 15 minutes after adding stop solution. IVD wef € € [ECIREP all SIA BIO-MEDICALS LTD. TRANSAS RinganWada, Daman - 396 210, India.