Uricostat enzimático AA
Contents
1. cccccsseseeeeeeeeaeeeeeeeeees 6 umol l DR R A A 2 0 mmol l NON PROVIDED REAGENTS Wiener lab s Calibrador A plus INSTRUCTIONS FOR USE Standard ready to use Reagents A and B ready to use They can be used sepa rately or as a Monoreagent mixing 4 parts of Reagent A 1 part of Reagent B e g 4 ml Reagent A 1 ml Reagent B WARNINGS Reagents are for in vitro diagnostic use Do not ingest Non provided with all kit sizes Avoid the contact with skin and eyes If spilt thoroughly wash affected area with water Use the reagents according to the working procedures for clinical laboratories The reagents and samples should be discarded according to the local regulations in force STABILITY AND STORAGE INSTRUCTIONS Provided Reagents stable in refrigerator 2 10 C until the expiration date printed of label While in use do not keep without refrigeration for extended periods of time Avoid contamination Monoreagent pre mixed in refrigerator 2 10 C is stable for 1 month since preparation date INSTABILITY OR DETERIORATION OF REAGENTS Failure to recover control values within the assigned range Standatrol S E 2 niveles could indicate deterioration and the Reagents should not be used Turbidity indicates Reagents deterioration Do not use Blank absorbance reading exceeding 0 200 O D or Stan dard readings abnormally low may indicate deterioration and the Reagents should not be used SAMPLE2. Serum plasma or urine a Collection obtain serum or plasma as usual Remove serum from clot as soon as possible within two hours from collection If urine is used it should be preferably fresh b Additives when using plasma use only heparin based anticoagulants c Known interfering substances Strongly reducing substances such as ascorbic acid vita min C Buscapina butyl hyoscine bromide interfere with the test Therefore therapy with ascorbic acid should be discontinued 24 hours before sample collection whenever possible No interference was observed from bilirubin up to 10 mg dl 100 mg l triglycerides up to 490 mg dl 4 9 g l hemoglo bin up to 180 mg dl and heparin up to 100 U ml See Young D S in References for effect of drugs on the present method d Stability and storage instructions samples should be preferably fresh If assay cannot be immediately performed serum or plasma samples may be stored for up to 3 days at 20 25 C 7 days at 2 10 C or 6 months at 20 C without preservatives Urine samples may be stored at pH gt 8 for up to 4 days at 20 25 C Do not refrigerate or freeze 864127522 01 p 7 9 REQUIRED MATERIAL non provided Spectrophotometer or photocolorimeter Adequate volumetric material Tubes or spectrophotometric square cuvettes Water bath at 37 C Watch or timer ASSAY CONDITIONS Wavelength 505 nm in spectrophotometer or in photocol orimeter with gre
3. C LIQUID LINE Uricostat enzimatico AA For acid uric determination in serum plasma or urine SUMMARY Uric acid is a metabolite found in purines nucleic acids and nucleoproteins Serum uric acid concentration usually varies from one indi vidual to another depending on several factors such as sex diet pattern ethnic origin genetic constitution pregnancy Abnormal levels of serum uric acid indicate metabolic disor ders of its precursors or inadequate excretion PRINCIPLE The analytical system is based on the following reaction UOD uric acid 2 H O O _ gt allantoin H O CO POD 2 H O 4 AP 3 5 DHS gt quinoneimine The amount of uric acid is determined by measuring the absorbance of this pigment UOD uricase POD peroxidase 4 AP 4 aminophenazone 3 5 DHS 3 5 dichlorohydroxybenzene sulfonic acid sodium salt PROVIDED REAGENTS S Standard 10 mg dl uric acid solution A Reagent A solution containing Good buffer pH 7 8 and 3 5 dichlorohydroxybenzene sulfonic acid sodium salt DHS B Reagent B solution containing Good buffer pH 7 8 4 ami nophenazone 4 AP uricase UOD peroxidase POD and potassium ferrocyanide Final concentrations Good Buffer ccc eztccienciuctstennnatietiendtonsictbtennansoxdieSenextdnes 50 mmol l LS E ee one eee ree ee ee ee eee gt 200 U I PD EE E E gt 1000 U I AAP E E E 0 10 mmol l Potassium ferrocyanide
4. en filter 490 530 nm Reaction temperature 37 C or 18 25 C Reaction time 5 minutes at 37 C or 20 minutes at 18 25 C Sample volume 20 ul Final reaction volume 1 02 ml Sample and Reagent volumes may be proportionally de creased or increased e g 50 ul Sample 2 5 ml monore agent or 10 ul Sample 500 ul monoreagent MANUAL PROCEDURE l TWO REAGENTS TECHNIQUE In three tubes or spectrophotometric cuvettes labeled B Blank S Reagent S or Calibrator and U Un known add B S U Standard 20 ul Sample 20ul Reagent A 800 ul 800ul 800 ul Reagent B 200 ul 200ul 200 ul Mix gently and incubate for 5 minutes in water bath at 37 C or for 20 minutes at room temperature 18 25 C Remove from bath Read in spectrophotometer at 505 nm or in photocolorimeter with green filter 490 530 nm setting the instrument to zero O D with the Blank Il MONOREAGENT TECHNIQUE Follow steps as described in Technique I using 1 ml of Monoreagent prepared following the Instructions for use Ill URINE TECHNIQUE Follow the above technique I or Il diluting the sample 1 10 with water or saline Calculate the results multi plying by the dilution factor used STABILITY OF FINAL REACTION Final reaction color is stable for 30 minutes thus absorbance should be read within that period CALCULATIONS 10 mg dl S M When Calibrador A plus is used see the uric acid con centration in its package insert ur
5. ficient between manual and automated methods was r 0 9971 slope b 0 9893 intercept a 0 2792 PARAMETERS FOR AUTOANALYZER For programming instructions check the user s manual of the autoanalyzer in use For calibration it must be used a serum based calibrator Wiener lab s Calibrador A plus WIENER LAB PROVIDES 225 ml 3 x 60 ml Reagent A 3 x 15 ml Reagent B Standard not included C d 1009320 225 ml 3 x 60 ml Reagent A 3 x 15 ml Reagent B Standard not included C d 1009635 250 ml 2 x 100 ml Reagent A 1 x 50 ml Reagent B Standard included Cod 1840107 400 ml 8 x 40 ml Reagent A 4 x 20 ml Reagent B Standard not included C d 1009277 500 ml 4 x 100 ml Reagent A 1 x 100 ml Reagent B Standard included C d 1840110 REFERENCES International Federation of Clinical Chemistry Clin Chim Acta 87 3 459 F 1978 Trinder P Ann Clin Biochem 6 24 1969 Young D S Effects of Drugs in Clinical Laboratory Tests 3 Ed AACC Press Washington DC 1990 NCCLS document Evaluation of the Linearity of Quantita tive Analytical Methods EP6 P 1986 NCCLS document Evaluation of Precision Performance EP5 A 1999 Tietz Fundamentals of clinical chemistry Burtis C Ash wood E 5 Edition WB Saunders 2001 864127522 01 p 9 9 SYMBOLS The following symbols are used in packaging for Wiener lab diagnostic reagents kits This product fulfi
6. ic acid mg l U xf where f 864127522 01 p 8 9 U absorbance reading of the unknown S absorbance reading of the Standard or Calibrator Example U 0 134 S 0 284 Uric acid in the Standard 10 mg dl f 10 mg dl 0 284 35 21 mg dl Uric acid in the sample 0 134 x 35 21 mg dl 4 72 mg dl QUALITY CONTROL METHOD Each time the test is performed analyze two levels of a qual ity control material Standatrol S E 2 niveles with known uric acid concentration REFERENCE VALUES Sera from 120 fasting individuals from both sexes with ages ranging from 20 to 45 years living in or near Rosario Argen tina with no symptoms of gout gouty nephropathy urate nephrolithiasis or other apparent disease were analyzed with Uricostat enzimatico AA liquida The central 95 of the results covers the following range Men 2 5 6 0 mg dl Women 2 0 5 0 mg dl In the literature Tietz N W the following reference value range is mentioned Serum or plasma Men 3 5 7 2 mg dl Women 2 6 6 0 mg dl Urine 250 a 750 mg 24 hours It is recommended that each laboratory establishes its own intervals and reference values taking into consideration age sex dietary habits and other factors SI SYSTEM UNITS CONVERSION Uric acid mg dl x 0 059 Uric acid mmol l Uric acid mg 24 hs x 0 0059 Uric acid mmol 24 hs PROCEDURE LIMITATIONS See Known interfering substances under SAMPLE PERFORMANCE The assays were performed
7. in a Express plus analyzer If using the kit with manual procedure user must validate that similar performance to that stated below is obtained a Reproducibility precision studies were performed ac cording to the guidelines contained in CLSI ex NCCLS document EP5 A and the following values were obtained Intra assay Level S D C V 3 39 mg dl 0 075 mg dl 2 21 5 36 mg dl 0 071 mg dl 1 32 Inter assay Level S D C V 3 39 mg dl 0 097 mg dl 2 86 5 36 mg dl 0 102 mg dl 1 90 TM Ciba Corning Diagnostics b Sensitivity based on an instrument minimal reading of 0 001 O D minimum detectable change in concentration under those conditions will be of approximately 0 03 mg dl c Linearity linearity studies were performed following the guidelines contained in CLSI ex NCCLS document EP6 P The reaction is linear up to 20 mg dl For higher values repeat the determination using half sample volume and multiply final result by 2 d Correlation Serum and plasma uric acid values of 100 specimens were determined using Uricostat enzimatico AA liquida and Uricostat enzimatico AA The correlation coefficient was r 0 9971 slope b 1 0167 and intercept a 0 2225 Manual vs automated procedures uric acid values of 30 samples were determined using the Uricostat enzimatico AA liquida kit with both manual and automated methods Sample uric acid concentrations covered a range from 1 7 to 18 2 mg dl Correlation coef
8. lls the requirements of the European Directive 98 79 EC for in vitro diagnostic medical devices Authorized representative in the European Community In vitro diagnostic medical device D Contains sufficient for lt n gt tests z Use by 9 Temperature limitation store at XK Do not freeze amp Biological risks gt Volume after reconstitution Cont Contents LOT Batch code ul Manufactured by xl Harmful a Corrosive Caustic x Irritant Ui Consult instructions for use Calibr Calibrator CONTROL Control gonTROL Positive Control Sous T Negative Control REF Catalog number wl Wiener Laboratorios S A 1 C Riobamba 2944 2000 Rosario Argentina A AAN E ota a ir T c Viviana E C tola W b Bioquimica lener a A N M A T Registered product Cert N 3464 99 2000 Rosario Argentina UR120903
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