testosterone ria ct ref ks24ct - Radim S.p.A.

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− Avoid splashing and aerosol formation; in such cases, carefully wash with a 3% sodium hypochlorite solution. Any cleaning material used for that purpose must be treated as potentially infectious and disposed accordingly. − The sodium azide used as preservative in some reagents may react with lead and copper plumbing; to prevent build-up of explosive metal azides, the reagents should be discarded by flushing the drain with large amounts of water. − According to Italian decree D.L. no. 22 dated 05.02.97, in compliance with EEC directives (91/156/EEC, 91/689/EEC, 94/62/EEC), all waste products originating from either manual and/or automated processing are classified as hazardous special waste material (European classification code 180103, infectious waste or potentially infectious waste). As such, they must be eliminated (by) delegating them to special enterprises (companies), qualified for waste collection and disposal. − All radioactive waste originating from either manual and/or automated processing, must be eliminated delegating them to special enterprises, qualified for waste collection and disposal, according to D.Lgs.241/2000. − Acquisition, storage, use and disposal of radioactive material (liquid and solid) are subjected to regulation and ordination of local authorities. � Radioactivity contents to the reference date is shown on the kit external label. SPECIMEN COLLECTION AND PREPARATION The assay can be performed in serum samples. Highly lipemic or hemolyzed samples must be discarded. Keep samples at 2-8°C for 1-2 days; for longer periods it is advisable to freeze samples at - 20°C. Make sure that samples are always perfectly clear before testing. Repeated freezing and thawing of samples should be avoided. ASSAY PROCEDURE − Allow reagents and samples to warm up at room temperature. − Mix samples by inversion before use. 1 - Prepare coated tubes for: Zero Calibrator (Bo), Calibrators (1-5), Control Serum and Samples. Use uncoated tubes for Total Activity (T) and Non-specific Binding (NSB). 2 - Pipette 100 µl of each Calibrator, Control Serum and Sample into the corresponding tubes. 3 - Pipette 100 µl of the Zero Calibrator into the Non-specific Binding (NSB) tubes. 4 - Add 500 µl of Radioactive Conjugate into all tubes. 5 - Mix on vortex. 6 - Incubate for 60 minutes at 37°C. 7 - Carefully aspirate the incubation mixture from all tubes, except those for total activity, with a vacuum pump or decant by drying the edges of the tubes with blot-paper. 8 - Count the radioactivity in the tubes for 1 minute by using a gamma counter. We suggest to check the background of the instrument before counting the assay. In order to avoid variations in the sensitivity of the system, the background should be reduced to a minimum or adjusted properly. ASSAY SCHEME: see p. 31 CALCULATION OF RESULTS Bo Cpm - NSB Cpm Binding Capacity % = ------------------------------- x 100 T Cpm - NSB Cpm Percent binding for calibrators, controls and samples = B Calibrators or Samples Cpm - NSB Cpm -----% = ----------------------------------------------------- x 100 Bo Bo Cpm - NSB Cpm Draw a calibration curve on logit-log or semi-log graph paper, by plotting the B/Bo% of each calibrator (yaxis) against the relative concentration (x-axis). Calculate the B/Bo% for each sample and read the concentration by interpolating on the calibration curve in order to obtain the Testosterone concentration in the tested samples, expressed in ng/mL. EXAMPLE OF CALCULATION KS24CT TESTOSTERONE RIA CT M104 - Rev.10 – 09/2007 - Pag. 10/32
The values shown below must be considered as an example and must not be used in place of experimental data. Description cpm B/Bo% B/T % Testosterone Total Activity (T) 38475 Non-specific binding (NSB) 160 0.41 Calibrator 0.0 ng/mL 15788 41.2 Calibrator 0.25 ng/mL 12734 80.6 Calibrator 0.5 ng/mL 11367 71.9 Calibrator 1.5 ng/mL 8729 55.2 Calibrator 6.0 ng/mL 4924 30.5 Calibrator 15.0 ng/mL 3061 19.3 Sample 9788 62.0 1.0 ng/mL NORMAL VALUES The values reported below are indicative. We suggest that each laboratory establishes its own normal range. Women 0.1 - 1.0 ng/mL Men 2.5 - 8.5 ng/mL Children 0.0 - 0.3 ng/mL Conversion: 1 ng/mL = 3.467 nmol/L. VALIDATION CRITERIA Before proceeding to calculation of results, make sure the control serum concentration is included within the value described on the Quality Control Sheet. PERFORMANCES OF THE ASSAY SPECIFICITY The present method has shown the following cross-reactions: 100% with Testosterone, 5.6% with 5-α- Dihydrotestosterone, 3.8% with 1, (5α)-Androsten-17 β-ol-3one, 2.6% with 19-Hydroxy- Androstenedione, 1.6% with Androstenedione and lower than 0.1% with Androstenediol, SHBG, Danazol, Estrone, DHEA-S, Estradiol, 5-α-Androstane-3 α -ol-17one and 5-β -Androstane- 17 α -ol 3one. The percentage of cross-reactivity is calculated by using the Abraham formula: X/Y x 100, X and Y being the weight of the substance to be tested and the weight of the cross-reacting substance respectively, both able to reduce the binding capacity by 50%. SENSITIVITY The sensitivity is the lowest dose of Testosterone capable of reducing the initial binding capacity by 5%. This dose is 0.017 ng/mL. PRECISION Precision was evaluated upon intra- and inter-assay variability, in 3 sera at different Testosterone concentrations. Intra-assay (Repeatability) Serum Mean ± S.D. C.V. Replicates (ng/mL) % no. a 0.53 ± 0.0327 6.1 10 b 0.91 ± 0.0356 3.9 10 c 4.45 ± 0.0685 1.5 10 Inter-assay (Reproducibility) Serum Mean ± S.D. C.V. Assays KS24CT TESTOSTERONE RIA CT M104 - Rev.10 – 09/2007 - Pag. 11/32
Page 1 and 2: TESTOSTERONE REF KS24CT Italiano p.
Page 3 and 4: DOSAGGIO RADIOIMMUNOLOGICO PER LA D
Page 5 and 6: − I materiali di origine umana ut
Page 7 and 8: Siero Media ± D.S. C.V. Replicati
Page 9: − Vortex mixer. − Water-bath, a
Page 13 and 14: RADIO-IMMUNASSAY ZUR QUANTITATIVEN
Page 15 and 16: − Entsprechend der italienischen
Page 17 and 18: (ng/mL) % Nr. a 0.53 ± 0.0327 6.1
Page 19 and 20: N.B.: vérifier toujours l’exact
Page 21 and 22: échantillon et par interpolation s
Page 23 and 24: RADIOINMUNOENSAYO PARA LA DETERMINA
Page 25 and 26: − Es necesario eliminar inmediata
Page 27 and 28: Repetibilidad (intra-ensayo) Suero
Page 29 and 30: RCNS H2O Ricostituire con / reconst
Page 31 and 32: Schema del Dosaggio, Assay Scheme,

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