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Experimental determination of the operative parameters for a double antibody solid phase radioimmunoassay of luteinizing and follicle-stimulating hormones in serum

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Ricerca in clinica e in laboratorio

Summary

The experimental parameters of a Double Antibody Solid Phase (DASP) radioimmunoassay (RIA) for the determination of serum luteinizing (LH) and follicle-stimulating (FSH) hormones, are described. The lowest detection limits can be fixed at about 20 pg LH and about 10 pg FSH. Evidence of parallelism among the dose-response curves and the dilution curves of different sera is provided for FSH-RIA and for about 92 % of the tested sera for LH-RIA. The antisera dilution curves are reported (final titres: 1/160·103 for anti-LH and 1/40·103 for anti-FSH). The traditional cross-reaction study in buffer indicates that 5.38 and 46.81 µU TSH/ml mimic 1 ng LH/ml and 1 ng FSH/ml respectively. The regression-line equations of antigen recovery (f=found, e=expected) are: LH f=0.94 LH e+0.91 and FSH f=1.05 FSH e+0.17; the same equations when TSH is added become: LH f=1.30 LH e+1.46 and FSH f=1.40 FSH e—0.37. It is concluded that LH-RIA is interfered by TSH over 8–10 µU TSH/ml and in function of TSH concentration; FSH-RIA is not interfered by TSH up to 4–5 ng FSH/ml; for higher FSH levels, TSH interferes according to a linear law (angular coefficient 1.40) and not in function of its concentration. LH and FSH levels throughout normal menstrual cycle in follicular phase, centre peak and luteal phase are: 3.14±1.16, 16.71±12.99, 2.31±1.04 ng LH/ml (mean±2SD) and 3.57±0.95, 5.72±2.90, 2.57±0.80 ng FSH/ml (mean±2SD) respectively. Finally, the within-assay coefficient of variation is 7.65 % for LH-RIA and 3.16 % for FSH-RIA.

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References

  1. Dotti C.: Diagnostica radioimmunologica. Modalità operative per la determinazione dei valori basali e per l’effettuazione degli stimoli farmacodinamici — La Ricerca Clin. Lab.3, 457, 1973.

    Google Scholar 

  2. Dotti C.: Gonadotropine: LH, FSH, HCG — In: Proceedings of the Convegno sul significato clinico del dosaggio radioimmunologico degli ormoni. Ravenna, Italy, May 11, 1974. (In Press).

  3. Dotti C.: Radioimmunoassay of Serum Triiodothyronine: Experimental Determination of the Operative Parameters and Clinical Results — J. nucl. Biol. Med.18, 6, 1974.

    Google Scholar 

  4. Dotti C.: Determinazione dei parametri operativi del dosaggio radioimmunologico della triiodotironina e del TSH — In: Proceedings of the Simposio sulle applicazioni cliniche del dosaggio radioimmunologico. Ferrara, Italy, October 3, 1974. (In Press).

  5. Dotti C., Portioli I.: Il dosaggio radioimmunologico dell’ormone tireotropo umano. Metodi di determinazione sperimentale dei parametri operativi — La Ricerca Clin. Lab.4, 251, 1974.

    Google Scholar 

  6. Feldmann H., Rodbard D.: Mathematical Theory of Radioimmunoassay — In:Odell W. D., Daughaday W. H. (Eds): Principles of Competitive Protein Binding Assays. J. B. Lippincott Co., Philadelphia, 1971; p. 158.

    Google Scholar 

  7. Greenwood F. C., Hunter W. M., Glover J. S.: The Preparation of I-131-Labelled Growth Hormone of High Specific Activity — Biochem. J.89, 114, 1963.

    PubMed  CAS  Google Scholar 

  8. Lee C. Y., Coulam C. B., Jiang N. S., Ryan R. J.: Receptors for Human Luteinizing Hormone in HumanCorpora lutea Tissue — J. clin. Endocr.36, 148, 1973.

    Article  PubMed  CAS  Google Scholar 

  9. Lemarchand-Béraud Th.: Specific Radioimmunoassay Procedures: Gonadotrophins — In: Intern. Training Course on RIA Techn. Pisa, Italy, May 3–14, 1971; SC/217, ITA.

  10. Reuter A. M., Gaspard U., Franchimont P.: Étude comparative des taux normaux des gonadotrophines suivant les méthodes radioimmunologiques employées — Institut National des radioéléments (IRE), B, February 4, 1973.

  11. Reuter A. M., Hendrick J.-C., Becker H., Franchimont P.: Comparaison de différentes méthodes de dosage radioimmunologique des gonadotrophines — In: Proceedings of the Symposium on Radioimmunoassay and Related Procedures in Clinical Medicine and Research. Istanbul, September 10–14, 1973; IAEA, SM-177/6.

  12. Reuter A. M., Hendrick J.-C., Franchimont P.: Mise au point d’un dosage radioimmunologique rapide des gonadotrophines — Ann. Biol. clin.31, 479, 1973.

    CAS  Google Scholar 

  13. Reuter A. M., Hendrick J.-C., Sulon J., Franchimont P.: Interference by Serum Protein with LH Radioimmunoassay Using Immunosorbant — Acta endocr. (Kbh.)72, 235, 1973.

    CAS  Google Scholar 

  14. Rodbard D.: Statistical Aspects of Radioimmunoassay — In:Odell W. D., Daughaday W. H. (Eds): Principles of Competitive Protein Binding Assays. J. B. Lippincott Co., Philadelphia, 1971; p. 204.

    Google Scholar 

  15. Spona J.: Rapid Assay for LH and Evaluation of Data by New Computer Program — In: Proceedings of the Symposium on Radioimmunoassay and Related Procedures in Clinical Medicine and Research. Istanbul, September 10–14, 1973; IAEA, SM-177/80.

  16. Wide L.: In:Diczfaluzy E. (Ed.): Immunoassay of Gonadotrophins — Acta endocr. (Kbh.)62 (Suppl. 142), 207, 1969.

    Google Scholar 

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Authors and Affiliations

  1. (1° Servizio Radiologico), Arcispedale S. Maria Nuova, Sezione di Medicina Nucleare, Viale Risorgimento 80, 42100, Reggio Emilia, Italia

    Claudio Dotti, Giordano Filippi, Claudio Castagnetti & Riccardo Franchini

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  1. Claudio Dotti
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  2. Giordano Filippi
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  3. Claudio Castagnetti
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  4. Riccardo Franchini
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Dotti, C., Filippi, G., Castagnetti, C. et al. Experimental determination of the operative parameters for a double antibody solid phase radioimmunoassay of luteinizing and follicle-stimulating hormones in serum. La Ricerca in Clin. Lab. 5, 249–261 (1975). https://doi.org/10.1007/BF02908288

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  • DOI: https://doi.org/10.1007/BF02908288

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