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Since the first radioimmunoassay (RIA) was developed in 1970s, many conventional RIAs and non-isotopic immunoassays (NIA) had been developed in which the degree of precision, accuracy, specificity and practicability have progressively increased. Recently ultrasensitive assay method at femtogram to determine testosterone in serum, saliva and feces is required for the study of sexual dysfunctions in male and female, monitoring the psychological stress and conditions, aging process such as menopause and partial androgen deficiency in aging male, the hormonal changes of small experimental animals etc. This review discussed the recentd evelopments of steroid assay methods, based upon the testosterone assay results of authors far 20 years, and the problems associated the assay set-up, the characterizations and applications of the established procedures, and desifls of assay, reliablity criteria, and the practical aspects of assay set-up and application, based upon the data of the authors. The present study demonstrates the general problems methods to be consider in order to set up the highly sensitive assay methods and to increase the assay quality and the necessity of assay quality control program. To improve the assay quality of each laboratory and to compare the assay results in homeland, the national QC programs should be organized.
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