Volume 2, Issue 1 (7-2004)
IJRM 2004, 2(1): 23-28 |
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Abstract: (1972 Views)
Background: Enzyme Linked ImmunoSorbent Assay (ELISA) has been described as an alternative to radioimmunoassay for the mammalian and nonmammalian steroids detection. In this study, a simple and rapid ELISA is described and validated for 4-pregnen-3,20, dione (progesterone). Materials and Methods: A general procedure for preparation of the acetylcholinesterase labelled steroid is described which is applicable to any steroid. Use of acetylcholinesterase tracer increased the sensitivity of assay so that reliable measurements of each steroid could be achieved with only 10 ?l of plasma. Results: Typical standard curves for progesterone steroids showed a workable range (detection limit) from 0.8 to 400 pg/well and the sensitivity of the assay taken as the concentration of steroid that induced 90% of B/B0, was 1.5 pg. Inter-assay variations that gave approximately 50% displacement was 9.2% for 10 replicates and intra-assay co-efficient of variation was less than 10% over the central part of the standard curve between 3 and 200 pg/well. There was a strong positive correlation (r>0.999) between the amount of steroid added to plasma and the amount measured. Conclusion: Method described here was applied to measure progesterone in plasma and this methodology could be of great interest to researchers measuring steroid hormones.
Type of Study: Original Article |
References
1. Basu A., and Shrivastav T.G. (2000) One step enzyme linked immunosorbent assay for direct estimation of serum cortisol. J Immunoassay 2: 39-50. [DOI:10.1080/01971520009349498] [PMID]
2. Canario A.V.M., and Scott A.P. (1991) Levels of 17 alpha, 20 alpha-dihydroxy-4-pregnen-3-one, 3 beta, 17 alpha,20-trihydroxy-5-BETA-pregnane, and other sex steroids, in blood plasma of male dab, Limanda limanda (marine flatfish) injected with human chorionic gonadotropin. General And Comparative Endocrinology 83: 258-264. [DOI:10.1016/0016-6480(91)90029-6]
3. Chen L., Ren W.H., Zhu S.L., Gao W., Zhou J., Jiang Y.Z., and Gu Y (2002) Effects of chronic cadmium loading on the testis and endocrine function of reproduction in male rats. 54: 258-262.
4. Christenson K.L., and Devoto L. (2003) Cholesterol transport and steroidogenesis by the corpus luteum. Reprod Biol Endocrinol. 1: 90 [DOI:10.1186/1477-7827-1-90] [PMID] [PMCID]
5. Cuisset B., Pradelles P., Kime D.E., Kuehn E.R., Babin P., Davail S., and Le Menn F. (1994) Enzyme immunoassay for 11-ketotestosterone using acetylcholinesterase as label: Application to the measurement of 11-ketotestosterone in plasma of Siberian sturgeon. Comparative Biochemistry and Physiology C Pharmacology Toxicology and Endocrinology 108: 229-241. [DOI:10.1016/1367-8280(94)90035-3]
6. Diamandis E.P., and Christopoulos T.K. (1996) Immunoassay. Academic Press, San Diego.
7. Dunlap K.D. (2002) Hormonal and body size correlates of electrocommunication behavior during dyadic interactions in a weakly electric fish, Apteronotus leptorhynchus. Horm Behav 41: 187-194. [DOI:10.1006/hbeh.2001.1744] [PMID]
8. Idler D.R. (1972) Steroids in nonmammalian vertebrates. Academic Press New York and London.
9. Kime D.E. (1993) Classical and non-clssical reproductive steroids in fish. Reviews in Fish Biology and Fisheries 3: 160-180. [DOI:10.1007/BF00045230]
10. Kime D.E., and Hews E.A. (1982) The effect of temperature on steroid biosynthesis by testes of the dogfish, Scyliorhinus caniculus. Comparative Biochemistry and Physiology. B: Comparative Biochemistry 71: 675-679. [DOI:10.1016/0305-0491(82)90480-1]
11. Kumari G.L., and Dhir R.N. (2003) Comparative studies with penicillinase, horseradish peroxidase, and alkaline phosphatase as enzyme labels in developing enzyme immunoassay of cortisol. J Immunoassay Immunochem 24: 173-190. [DOI:10.1081/IAS-120020083] [PMID]
12. Larsson D.G., Mayer I., Hyllner S.J., and Forlin L. (2002) Seasonal variations of vitelline envelope proteins, vitellogenin, and sex steroids in male and female eelpout (Zoarces viviparus). Gen Comp Endocrinol 125: 184-96. [DOI:10.1006/gcen.2001.7740] [PMID]
13. Manire C.A., Rasmussen L.E., and Gross T.S. (1999) Serum steroid hormones including 11-ketotestosterone, 11-ketoandrostenedione, and dihydroprogesterone in juvenile and adult bonnethead sharks, Sphyrna tiburo. J Exp Zool 284: 595-603.
https://doi.org/10.1002/(SICI)1097-010X(19991001)284:5<595::AID-JEZ15>3.0.CO;2-6 [DOI:10.1002/(SICI)1097-010X(19991001)284:53.0.CO;2-6]
14. Mayer I., Rand-Weaver M., and Borg B. (1998) Effects of gonadectomy and steroids on plasma and pituitary levels of somatolactin in Atlantic salmon, Salmo salar. Gen Comp Endocrinol 109: 223-231. [DOI:10.1006/gcen.1997.7015] [PMID]
15. Melamed P., Gur G., Rosenfeld H., Elizur A., Schulz R.W., and Yaron Z. (2000) Reproductive development of male and female tilapia hybrids (Oreochromis niloticus x O. aureus) and changes in mRNA levels of gonadotropin (GtH) Ibeta and IIbeta subunits. J Exp Zool 286: 64-75.
https://doi.org/10.1002/(SICI)1097-010X(20000101)286:1<64::AID-JEZ6>3.0.CO;2-D [DOI:10.1002/(SICI)1097-010X(20000101)286:13.0.CO;2-D]
16. Nishi K., Chikae M., Hatano Y., Mizukami H., Yamashita M., Sakakibara R., and Tamiya E. (2002) Development and application of a monoclonal antibody-based sandwich ELISA for quantification of Japanese medaka (Oryzias latipes) vitellogenin. Comp Biochem Physiol C Toxicol Pharmacol 132: 161-169. [DOI:10.1016/S1532-0456(02)00058-3]
17. Pall M.K, Mayer I., and Borg B. (2002) Androgen and Behavior in the Male Three-Spined Stickleback, Gasterosteus aculeatus. II. Castration and 11-Ketoandrostenedione Effects on Courtship and Parental Care during the Nesting Cycle. Horm Behav 42: 337-344. [DOI:10.1006/hbeh.2002.1820] [PMID]
18. Pandey P.K., Shrivastav T.G., Kumari G.L., Rao P.N., Grover P.K., and Murthy H.G. (1990) Enzyme immunosorbant assay of oestradiol in unextracted plasma using penicillinase as label. Clin Chim Acta 190: 175-184. [DOI:10.1016/0009-8981(90)90171-N]
19. Pradelles P., Grassi J., and Maclouf J. (1985) Enzyme immunoassays of eicosanoids using acetylcholinesterase as label: an alternative to radioimmunoassay. Analyt. Chem. 57: 1170-1173. [DOI:10.1021/ac00284a003] [PMID]
20. Rohr D.H., Lokman P.M., Davie P.S., and Young G. (2001) 11-Ketotestosterone induces silvering-related changes in immature female short-finned eels, Anguilla australis. Comp Biochem Physiol A Mol Integr Physiol 130: 701-714. [DOI:10.1016/S1095-6433(01)00402-0]
21. Scott A.P., Sumpter J.P., et al. (1983) Hormone changes during ovulation in the rainbowe trout (Salmo gairdneri Richardson). General and Comparative Endocrinology 49: 128-134. [DOI:10.1016/0016-6480(83)90016-3]
22. Tattersfield L., Matthiessen P., Campbell P., Grandy N., and Lange R. (1997) SETAC-Europe/OECD/EC Expert Workshop on Endocrine Modulators and Wildlife: Assessment and Testing (EMWAT). Proceeding of SETAC-Europe/OECD/EC Expert Workshop, Veldhoven, The Netherlands, 10-13th April 1997. SETAC-Europe, Amsterdam. 1.
23. Tomita Y., Engelman R.W., Bauer-Sardina I., Day N.K., and Good R.A. (1992) Improved enzyme immunosorbent assay for mouse prolactin using penicillinase as label. J Immunol Methods 151: 269-275. [DOI:10.1016/0022-1759(92)90127-F]
24. Wade M.G., Foster W.G., Younglai E.V., McMahon A., Leingartner K., Yagminas A., Blakey D., Fournier M, Desaulniers D., and Hughes C.L. (2002) Effects of subchronic exposure to a complex mixture of persistent contaminants in male rats: systemic, immune, and reproductive effects. Toxicological Sciences: An Official Journal of the Society of Toxicology 67: 131-143. [DOI:10.1093/toxsci/67.1.131] [PMID]
25. Weltzien F.A., Taranger G.L., Karlsen O., and Norberg B. (2002) Spermatogenesis and related plasma androgen levels in Atlantic halibut (Hippoglossus hippoglossus L.). Comp Biochem Physiol A Mol Integr Physiol 132: 567-575. [DOI:10.1016/S1095-6433(02)00092-2]
26. Yamada H., Satoh R., Ogoh M., Takaji K., Fujimoto Y., Hakuba T., Chiba H., Kambegawa A., and Iwata M. (2002) Circadian changes in serum concentrations of steroids in Japanese char Salvelinus leucomaenis at the stage of final maturation. Zoolog Sci 19: 891-898. [DOI:10.2108/zsj.19.891] [PMID]
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