RAT FSH IRMA C.T. (AH R004) Package insert / Instruction for use
RAT FSH IRMA C.T. (AH R004) Package insert / Instruction for use
RAT FSH IRMA C.T. (AH R004) Package insert / Instruction for use
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<strong>RAT</strong> <strong>FSH</strong> <strong>IRMA</strong> C.T. (<strong>AH</strong> <strong>R004</strong>) <strong>Package</strong> <strong>insert</strong> / <strong>Instruction</strong> <strong>for</strong> <strong>use</strong><br />
Immunoradiometric assay on coated tubes <strong>for</strong> the determination of Follitropin in rat serum<br />
or plasma<br />
1. Intended <strong>use</strong><br />
For IN VITRO determination of serum and plasma rat follitropin (r<strong>FSH</strong>) levels.<br />
2. Principle of the method<br />
The rat <strong>FSH</strong> <strong>IRMA</strong> is a one step solid phase immunoradiometric assay which<br />
offers high affinity and specificity <strong>for</strong> two different epitopes on <strong>FSH</strong>. A first<br />
monoclonal anti-<strong>FSH</strong> antibody bound to a polystyrene tube will capture the<br />
<strong>FSH</strong> of the sample in the presence of a second 125 I labelled monoclonal anti-<br />
<strong>FSH</strong> antibody.<br />
Following the incubation and the one step <strong>for</strong>mation of the solid phase-<strong>FSH</strong>-<br />
125<br />
I labelled monoclonal antibody sandwich, the tube is washed to remove<br />
excess of unbound labelled antibody. The radioactivity of the sandwich is<br />
directly proportional to the amount of <strong>FSH</strong> present in the sample.<br />
Rat samples concentrations are read from a calibration curve and the results<br />
are expressed in ng/ml.<br />
3. Warnings and precautions<br />
For in vitro diagnostic <strong>use</strong><br />
It must be handled by specialized staff.<br />
Good laboratory and safety practices are advisable.<br />
Warning<br />
This kit contains 125 I emitting X and γ ionizing rays.<br />
This radioactive material may be received, acquired, possessed and <strong>use</strong>d only<br />
by persons in clinical or hospital laboratories who are authorized by competent<br />
authorities and only <strong>for</strong> in vitro clinical or laboratory tests not involving internal<br />
or external administration of the material, or the radiation therefrom , to human<br />
beings or animals. Its receipt, acquisition, possession, <strong>use</strong>, transfer, the waste<br />
disposal and the people protection are subject to the State and local<br />
regulations.<br />
Use impermeable gloves and appropriate protection clothes.<br />
Warning : Some components contain sodium azide ( 98 % m/m).<br />
Seeing the lack of international references <strong>for</strong> r<strong>FSH</strong>, we have established a<br />
QC procedure which guarantees batch to batch reproducibility <strong>for</strong> standard<br />
curve calibration.<br />
4.5. CONTROL<br />
1 vials lyophilized rat <strong>FSH</strong> supplied in horse serum containing preservatives<br />
(NaN3 < 1g/l). The control has to be assayed along with the samples and<br />
the result compared to the range printed on the vial label.<br />
4.6. BUF WASH 10x<br />
1 bottle (100 ml) of concentrated buffered solution with preservatives<br />
(NaN3
6.4. Example of a typical assay<br />
Contents<br />
(ng/ml)<br />
cpm 1st<br />
duplicate<br />
cpm 2nd<br />
duplicate<br />
Mean<br />
count rate<br />
B/T %<br />
"Total counts" - 115829 115391 115610 - -<br />
CAL 0 0 178 171 174 0.15 -<br />
CAL 1 2 1337 1283 1310 1.13<br />
CAL 2 10 5436 5420 5428 4.70<br />
CAL 3 20 10295 10000 10148 8.78<br />
CAL 4 50 21691 20963 21327 18.45<br />
CAL 5 100 31748 33909 32828 28.40<br />
CAL 6 200 48581 46103 47342 40.95<br />
r<strong>FSH</strong><br />
conc.<br />
(ng/ml)<br />
CONTROL 11.3-16.7 7575 7542 7559 6.54 14.4<br />
Sample 1 4746 4722 4734 4.09 8.6<br />
Sample 2 16350 15900 16125 13.95 34.3<br />
Sample 3 29025 28842 28933 25.03 84.6<br />
Example of a typical assay, do not <strong>use</strong> <strong>for</strong> calculations<br />
7. Expected normal values<br />
It is recommended that each laboratory establishes its own reference values.<br />
Concentrations range <strong>for</strong> normal male subjects from 14,75 to 19,05 ng/ml and<br />
<strong>for</strong> normal female subjects from 5.16 to 10.44 ng/ml in our preliminary studies.<br />
8. Limitation of the procedure<br />
Do not <strong>use</strong> strongly lipemic, haemolyzed, icteric or turbid specimens<br />
9. Quality control<br />
Use the control provided <strong>for</strong> each assay.<br />
If, in normal using conditions, the control is out the acceptable range, the<br />
sample results can’t be validated. Please contact the manufacturer.<br />
10. Per<strong>for</strong>mance characteristics<br />
10.1. Specificity<br />
The assay was found to be specific <strong>for</strong> r-<strong>FSH</strong>. <strong>FSH</strong> from other species can<br />
crossreact partially in this assay. The weak cross-reactivities measured with<br />
other rat pituitary hormones are due to contamination of the hormonal<br />
preparation as checked by radioreceptor assays.<br />
10.2. Analytical sensitivity<br />
The minimum detectable concentration of rat <strong>FSH</strong> has been assayed at 0.2<br />
ng/ml and corresponds to the concentration given by two standard deviation<br />
above the mean cpm of 20 replicates determinations of the zero calibrator.<br />
10.3. Imprecision<br />
Repeatability (Within assay variation) and reproductibility (Between assay<br />
variation)<br />
Mean<br />
value<br />
(mIU/ml)<br />
Repeatability<br />
Within assay variation<br />
18 replicates (% CV)<br />
Reproducibility<br />
Between assay variation<br />
8 separate assays (% CV)<br />
Pool 1 15.2 2.9 7.6<br />
Pool 2 34.4 2.3 7.8<br />
Pool 3 80.3 2.4 6.4<br />
11. Bibliography<br />
1. J. CLOSSET, G. HENNEN<br />
Biopotency of highly purified <strong>FSH</strong> and LH on the testis of immature<br />
hypophysectomized rats.<br />
J Endocrinol, 119 : 542, 1988.<br />
2. K.J. TEERDS, J. CLOSSET, R.F.G. ROMMERTS, D.G. DE ROOIJ, D.M.<br />
STOCCO, B. COLENBRANDER, J. GWENSING, G. HENNEN<br />
Effects of pure <strong>FSH</strong> and LH preparations on the number and function of<br />
Leydig cells in immature hypophysectomized rats.<br />
J. Endocrinol, 120 : 97-106, 1989.<br />
BIOCODE –HYCEL<br />
Rue E. Solvay, 101-103 - 4000 Liège - Belgium<br />
Tel. **32/4/252.26.36 - Fax **32/4/252.51.96<br />
E-MAIL : ivd@biocode.be<br />
Web Site : http://www.biocodehycel.com<br />
Catalogue number <strong>AH</strong> <strong>R004</strong><br />
<strong>Package</strong> <strong>insert</strong> /date of issue : 2003-12-17