Instructions for Use
Testosterone RIA (CT)
Radioimmunoassay (Coated tube)
for the quantitative determination of
testosterone in human serum and plasma.
MG12191
96
2-8°C
I B L I N T E R N A T I O N A L G M B H
Flughafenstrasse 52a Phone: +49 (0)40-53 28 91-0 IBL@IBL-International.com
D-22335 Hamburg, Germany Fax: +49 (0)40-53 28 91-11 www.IBL-International.com
Read entire protocol before use.
TESTO-RIA-CT
I. INTENDED USE
Radioimmunoassay for the in vitro quantitative measurement of human Testosterone (TESTO) in
serum.
II. GENERAL INFORMATION
A. Proprietary name : TESTO-RIA-CT Kit
B. Catalog number : MG12191 : 96 tests
III. CLINICAL BACKGROUND
A. Biological activity
Testosterone is a C-19 steroid hormone (molecular weight: 288 Da) which is produced from
androstenedione in the testes, adrenals and ovaries. Testosterone is a precursor along with androstenedione
of the estrogen steroids.
B. Clinical applications
· Clinical significance of testosterone level : Source of testosterone :
. Women : Ovary, adrenal cortex, peripheral tissues (by conversion of 50-60% other steroids).
. Men : Testes > 90%, adrenal cortex, peripheral tissues.
· Clinical diseases with high level of testosterone :
. Women : Hirsutism and virilization, polycystic ovary syndrome, congenital adrenal hyperplasia (with
170H-PROG), tumors of adrenal and ovarian origin, breast cancer.
. Men : Disease of the hypothalamic pituitary unit, some malignant testicular tumors, congenital adrenal
hyperplasia, prostate cancer.
· Clinical diseases with low level of testosterone :
Primary or secondary hypogonadism, Klinefelter's syndrome, other chromosomal alteration,
hypopituitarism, enzymatic defects, orchidectomy and cryptorchidism, testicular feminization, hepatic
cirrhosis, some autoimmune diseases for example : Sjögren's syndrome, systemic lupus.
· Other domains for measurement of testosterone level :
. In vitro fertilization : the women with high respons to gonadotrophin have a significant increase in
testosterone.
. Parameter of the prepuberty and puberty.
. Determination of foetal sex in amniotic fluid.
. Free testosterone is significantly raised in both male and female acne sufferers.
. Follow up of cancer and in pathological situations; low testosterone syndrom.
IV. PRINCIPLES OF THE METHOD IX. SPECIMEN COLLECTION AND PREPARATION
A fixed amount of 125I labelled steroid competes with the steroid to be measured - Serum samples must be kept at 2-8°C.
present in the sample or in the calibrator for a fixed amount of antibody sites - If the test is not run within 24 hrs, storage at -20°C is recommended.
being immobilized to the wall of a polystyrene tube. Neither extraction nor - Avoid subsequent freeze-thaw cycles.
chromatography are required because of the high specificity of the coated
antibodies. After 3 hours incubation at 37°C, an aspiration step terminates the
competition reaction. The tubes are then washed with 3 ml of wash solution and X. PROCEDURE
aspirated again. A calibration curve is plotted and the testosterone concentrations
of the samples are determined by dose interpolation from the calibration curve. A. Handling notes
Do not use the kit or components beyond expiry date.
Do not mix materials from different kit lots.
V. REAGENTS PROVIDED Bring all the reagents to room temperature prior to use.
Thoroughly mix all reagents and samples by gentle agitation or swirling.
Use a clean disposable pipette tip for addition of each different reagent and
Reagents 96 Test Kit Colour Reconstitution
sample in order to avoid cross-contamination. High precision pipettes or
Code
automated pipetting equipment will improve the precision.
Respect the incubation times.
Tubes coated with anti 2 x 48 green Ready for use
Prepare a calibration curve for each run, do not use data from previous runs.
TESTO
B. Procedure
125 1 vial red Ready for use 1. Label coated tubes in duplicate for each calibrator, control and sample. For
Ag I
55 ml
the determination of total counts, label 2 normal tubes
180 kBq
TRACER: 125
Iodine labelled
2. Briefly vortex calibrators, controls and samples and dispense 50µl of each
TESTO (HPLC grade) in into the respective tubes.
phosphate-citrate buffer with 3. Dispense 500 µl of 125Iodine labelled TESTO into each tube, including the
bovine gelatin and azide ( ± SD CV Serum N ± SD CV
(ng/ml) (%) (ng/ml) (%)
Premenopausal women were with normal luteal phase (Progesterone > 30 nmol/l),
not on clomid and with no evidence of irregular cycle. Postmenopausal women
(age : 28 to 61) had FSH > 30 IU/l and most of these patients were routine
A 10 0.69 ± 0.03 4.6 A 20 0.55 ± 0.03 6.2
B 10 4.35 ± 0.14 3.3 B 20 3.51 ± 0.17 4.8 assessment of confirming recent post-menopausal status or premature ovarian
C 9 9.82 ± 0.44 4.4 failure.
Identification Range (*) Mean SD n
SD: Standard Deviation; CV: Coefficient of variation
(ng/ml)
D. Accuracy Females (determined in UK)
DILUTION TEST . Premenopausal 0.11 - 0.79 0.39 0.17 100
. Postmenopausal < 0.06 - 0.50 0.22 0.13 60
Sample Dilution Theoretical Measured
Concent. Concent. Males 1.34 – 6.25 3.89 1.46 71
(ng/ml) (ng/ml)
(*) The range is based on 2.5 % and 97.5 % percentiles
A 1/1 - 8.72
1/2 4.36 4.08
1/4 2.18 2.00
1/8 1.09 1.08
XVI. PRECAUTIONS AND WARNINGS
1/16 0.55 0.56
1/32 0.27 0.23 Safety
For in vitro diagnostic use only.
B 1/1 - 6.98 This kit contains 125I (half-life: 60 days), emitting ionizing X (28 keV) and γ (35.5
1/2 3.49 3.33 keV) radiations.
1/4 1.75 1.59 This radioactive product can be transferred to and used only by authorized
1/8 0.87 0.81
persons; purchase, storage, use and exchange of radioactive products are subject
1/16 0.44 0.42
1/32 0.22 0.18 to the legislation of the end user's country. In no case the product must be
administered to humans or animals.
All radioactive handling should be executed in a designated area. away from
Samples were diluted with zero calibrator. regular passage. A logbook for receipt and storage of radioactive materials must
be kept in the lab. Laboratory equipment and glassware, which could be
contaminated with radioactive substances, should be segregated to prevent cross
RECOVERY TEST contamination of different radioisotopes.
Sample added TESTO Recovered Recovered Any radioactive spills must be cleaned immediately in accordance with the
(ng/ml) TESTO (%) radiation safety procedures. The radioactive waste must be disposed of following
(ng/ml) the local regulations and guidelines of the authorities holding jurisdiction over the
laboratory. Adherence to the basic rules of radiation safety provides adequate
1 0.22 0.19 86.4% protection.
0.46 0.51 110.9% The human blood components included in this kit have been tested by European
1.36 1.5 110.3% approved and/or FDA approved methods and found negative for HbsAg, anti-
3.28 3.09 94.2%
HCV, anti-HIV-1 and 2. No known method can offer complete assurance that
9.8 12.2 124.5%
human blood derivatives will not transmit hepatitis, AIDS or other infections.
Therefore, handling of reagents, serum or plasma specimens should be in
accordance with local safety procedures.
Conversion factor :
All animal products and derivatives have been collected from healthy animals.
From ng/ml to nmol/L : x 3.47 Bovine components originate from countries where BSE has not been reported.
From nmol/L to ng/ml : x 0.288
Nevertheless, components containing animal substances should be treated as
potentially infectious.
The concentrations of the calibrators are determined with the ID-GC/MS Avoid any skin contact with reagents (sodium azide as preservative). Azide in
reference method.
this kit may react with lead and copper in the plumbing and in this way form
highly explosive metal azides. During the washing step, flush the drain with a
large amount of water to prevent azide build-up.
Do not smoke, drink, eat or apply cosmetics in the working area. Do not pipette
by mouth. Use protective clothing and disposable gloves.
8. R. MARUYAMA et al. (1987)
XVII. BIBLIOGRAPHY Sex-Steroid-Binding Plasma Protein (SBP), Testosterone, Oestradiol
and DHEA in Prepuberty and Puberty.
1. J.L. ANDREYKO et al. (1986) Acta Endocrinologica, 114: 60-67
Role of Serum Androgens and Sex Hormone Binding Globulin
Capacity in the Evaluation of Hirsutism in Women. 9. P. HOLDUNIA, C. WALTER et al (1992)
Clin. Biochem, Vol. 19, 58-61. A clinical evaluation of a direct radioimmunoassay of testosterone.
Clin. Chim. Acta, 214:31-43.
2. A. BIZZARO et al.
Influence of Testosterone Therapy on Clinical and Immunological
features of Autoimmune Diseases Associated with Klinefelter's XVIII. SUMMARY OF THE PROTOCOL
Syndrome
J. Clin. Endocrin. Metab. Vol. 64, N°1: 32-36
TOTAL CALIBRATORS SAMPLE (S)
3. M. CARRABBA et al (1985) COUNTS CONTROLS
Abnormalities of Sex hormones in Men with Systemic Lupus µl µl µl
Erythematosus.
Clin. Rheumatology, N° 4: 422-425. Calibrators (0 to 5) - 50 -
Samples, Controls - - 50
4. P. HILL et al. (1985) Tracer 500 500 500
Plasma Testosterone and Breast Cancer.
Eur.J Cancer Clin. Oncol, Vol. 21, N°10, pp. 1265-1266 Incubation 3 hours at 37°C
5. CG. MAHLCK et al (1986) /Separation - Aspirate (or decant)
Testosterone, SHBG and Albumin in Patients with ovarian carcinoma. Working Wash solution 3.0 ml
Acta Obstet. Gynecol. Scand. 65: 533-S38. Separation Aspirate (or decant)
6. D.M.D PERERA et 31 (1987)
Amniotic Fluid Testosterone and testosterone Glucuronide Levels in Counting Count tubes for 60 seconds
the determination of Foetal Sex.
J. Steroid Biochem., Vol. 26,N°2,pp.273-277.
7. J. TRACHTENBERG (1987)
IBL Catalogue Nr : P.I. Number : Revision nr :
Experimental Treatment of Prostatic Cancer by intermittent
MG12191 1700507/en 060206/1
Hormonal Therapy.
J. Urology, Vol. 137, pp. 785-788.
Revision date : 2006-02-06
Symbols / Symbole / Symbôles / Símbolos / Símbolos / Σύµβολα
REF Cat.-No.: / Kat.-Nr.: / No.- Cat.: / Cat.-No.: / N.º Cat.: / N.–Cat.: / Αριθµός-Κατ.:
LOT Lot-No.: / Chargen-Bez.: / No. Lot: / Lot-No.: / Lote N.º: / Lotto n.: / Αριθµός -Παραγωγή:
Use by: / Verwendbar bis: / Utiliser à: / Usado por: / Usar até: / Da utilizzare entro: /
Χρησιµοποιείται από:
No. of Tests: / Kitgröße: / Nb. de Tests: / No. de Determ.: / N.º de Testes: / Quantità dei tests: /
Αριθµός εξετάσεων:
CONC Concentrate / Konzentrat / Concentré / Concentrar / Concentrado / Concentrato / Συµπύκνωµα
LYO Lyophilized / Lyophilisat / Lyophilisé / Liofilizado / Liofilizado / Liofilizzato / Λυοφιλιασµένο
In Vitro Diagnostic Medical Device. / In-vitro-Diagnostikum. / Appareil Médical pour Diagnostics In
IVD Vitro. / Dispositivo Médico para Diagnóstico In Vitro. / Equipamento Médico de Diagnóstico In
Vitro. / Dispositivo Medico Diagnostico In vitro. / Ιατρική συσκευή για In-Vitro ∆ιάγνωση.
Evaluation kit. / Nur für Leistungsbewertungszwecke. / Kit pour évaluation. / Juego de Reactivos
para Evaluació. / Kit de avaliação. / Kit di evaluazione. / Κιτ Αξιολόγησης.
Read instructions before use. / Arbeitsanleitung lesen. / Lire la fiche technique avant emploi. /
Lea las instrucciones antes de usar. / Ler as instruções antes de usar. / Leggere le istruzioni
prima dell’uso. / ∆ιαβάστε τις οδηγίες πριν την χρήση.
Keep away from heat or direct sun light. / Vor Hitze und direkter Sonneneinstrahlung schützen. /
Garder à l’abri de la chaleur et de toute exposition lumineuse. / Manténgase alejado del calor o la
luz solar directa. / Manter longe do calor ou luz solar directa. / Non esporre ai raggi solari. / Να
φυλάσσεται µακριά από θερµότητα και άµεση επαφή µε το φως του ηλίου.
Store at: / Lagern bei: / Stocker à: / Almacene a: / Armazenar a: / Conservare a: / Αποθήκευση
στους:
Manufacturer: / Hersteller: / Fabricant: / Productor: / Fabricante: / Fabbricante: / Παραγωγός:
Caution! / Vorsicht! / Attention! / ¡Precaución! / Cuidado! / Attenzione! / Προσοχή!
Symbols of the kit components see MATERIALS SUPPLIED.
Die Symbole der Komponenten sind im Kapitel KOMPONENTEN DES KITS beschrieben.
Voir MATERIEL FOURNI pour les symbôles des composants du kit.
Símbolos de los componentes del juego de reactivos, vea MATERIALES SUMINISTRADOS.
Para símbolos dos componentes do kit ver MATERIAIS FORNECIDOS.
Per i simboli dei componenti del kit si veda COMPONENTI DEL KIT.
Για τα σύµβολα των συστατικών του κιτ συµβουλευτείτε το ΠΑΡΕΧΟΜΕΝΑ ΥΛΙΚΑ.
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LIABILITY: Complaints will only be accepted in written and if all details of the test performance and results are included (complaint form available
from IBL or supplier). Any modification of the test procedure or exchange or mixing of components of different lots could negatively affect the results.
These cases invalidate any claim for replacement. Regardless, in the event of any claim, the manufacturer’s liability is not to exceed the value of the
test kit. Any damage caused to the kit during transportation is not subject to the liability of the manufacturer.
Symbols Version 3.5 / 2008-10-01