RAT FSH IRMA C.T. (AH R004) Package insert / Instruction for use

RAT FSH IRMA C.T. (AH R004) Package insert / Instruction for use
Immunoradiometric assay on coated tubes for the determination of Follitropin in rat serum
or plasma
1. Intended use
For IN VITRO determination of serum and plasma rat follitropin (rFSH) levels.
2. Principle of the method
The rat FSH IRMA is a one step solid phase immunoradiometric assay which
offers high affinity and specificity for two different epitopes on FSH. A first
monoclonal anti-FSH antibody bound to a polystyrene tube will capture the
FSH of the sample in the presence of a second 125 I labelled monoclonal anti-
FSH antibody.
Following the incubation and the one step formation of the solid phase-FSH-
125
I labelled monoclonal antibody sandwich, the tube is washed to remove
excess of unbound labelled antibody. The radioactivity of the sandwich is
directly proportional to the amount of FSH present in the sample.
Rat samples concentrations are read from a calibration curve and the results
are expressed in ng/ml.
3. Warnings and precautions
For in vitro diagnostic use
It must be handled by specialized staff.
Good laboratory and safety practices are advisable.
Warning
This kit contains 125 I emitting X and γ ionizing rays.
This radioactive material may be received, acquired, possessed and used only
by persons in clinical or hospital laboratories who are authorized by competent
authorities and only for in vitro clinical or laboratory tests not involving internal
or external administration of the material, or the radiation therefrom , to human
beings or animals. Its receipt, acquisition, possession, use, transfer, the waste
disposal and the people protection are subject to the State and local
regulations.
Use impermeable gloves and appropriate protection clothes.
Warning : Some components contain sodium azide ( 98 % m/m).
Seeing the lack of international references for rFSH, we have established a
QC procedure which guarantees batch to batch reproducibility for standard
curve calibration.
4.5. CONTROL
1 vials lyophilized rat FSH supplied in horse serum containing preservatives
(NaN3 < 1g/l). The control has to be assayed along with the samples and
the result compared to the range printed on the vial label.
4.6. BUF WASH 10x
1 bottle (100 ml) of concentrated buffered solution with preservatives
(NaN3

6.4. Example of a typical assay
Contents
(ng/ml)
cpm 1st
duplicate
cpm 2nd
duplicate
Mean
count rate
B/T %
"Total counts" - 115829 115391 115610 - -
CAL 0 0 178 171 174 0.15 -
CAL 1 2 1337 1283 1310 1.13
CAL 2 10 5436 5420 5428 4.70
CAL 3 20 10295 10000 10148 8.78
CAL 4 50 21691 20963 21327 18.45
CAL 5 100 31748 33909 32828 28.40
CAL 6 200 48581 46103 47342 40.95
rFSH
conc.
(ng/ml)
CONTROL 11.3-16.7 7575 7542 7559 6.54 14.4
Sample 1 4746 4722 4734 4.09 8.6
Sample 2 16350 15900 16125 13.95 34.3
Sample 3 29025 28842 28933 25.03 84.6
Example of a typical assay, do not use for calculations
7. Expected normal values
It is recommended that each laboratory establishes its own reference values.
Concentrations range for normal male subjects from 14,75 to 19,05 ng/ml and
for normal female subjects from 5.16 to 10.44 ng/ml in our preliminary studies.
8. Limitation of the procedure
Do not use strongly lipemic, haemolyzed, icteric or turbid specimens
9. Quality control
Use the control provided for each assay.
If, in normal using conditions, the control is out the acceptable range, the
sample results can’t be validated. Please contact the manufacturer.
10. Performance characteristics
10.1. Specificity
The assay was found to be specific for r-FSH. FSH from other species can
crossreact partially in this assay. The weak cross-reactivities measured with
other rat pituitary hormones are due to contamination of the hormonal
preparation as checked by radioreceptor assays.
10.2. Analytical sensitivity
The minimum detectable concentration of rat FSH has been assayed at 0.2
ng/ml and corresponds to the concentration given by two standard deviation
above the mean cpm of 20 replicates determinations of the zero calibrator.
10.3. Imprecision
Repeatability (Within assay variation) and reproductibility (Between assay
variation)
Mean
value
(mIU/ml)
Repeatability
Within assay variation
18 replicates (% CV)
Reproducibility
Between assay variation
8 separate assays (% CV)
Pool 1 15.2 2.9 7.6
Pool 2 34.4 2.3 7.8
Pool 3 80.3 2.4 6.4
11. Bibliography
1. J. CLOSSET, G. HENNEN
Biopotency of highly purified FSH and LH on the testis of immature
hypophysectomized rats.
J Endocrinol, 119 : 542, 1988.
2. K.J. TEERDS, J. CLOSSET, R.F.G. ROMMERTS, D.G. DE ROOIJ, D.M.
STOCCO, B. COLENBRANDER, J. GWENSING, G. HENNEN
Effects of pure FSH and LH preparations on the number and function of
Leydig cells in immature hypophysectomized rats.
J. Endocrinol, 120 : 97-106, 1989.
BIOCODE –HYCEL
Rue E. Solvay, 101-103 - 4000 Liège - Belgium
Tel. **32/4/252.26.36 - Fax **32/4/252.51.96
E-MAIL : ivd@biocode.be
Web Site : http://www.biocodehycel.com
Catalogue number AH R004
Package insert /date of issue : 2003-12-17