title: A highly specific heterologous enzyme immunoassay for 5 alpha-androstane-3 alpha, 17 beta-diol 17-glucuronide (androstanediol-17G) and developmental patterns of urinary androstanediol-17G excretions.
abstract: We established a highly specific enzyme immunoassay (EIA) for 5 alpha-androstane-3 alpha, 17 beta-diol 17-glucuronide (androstanediol-17G). Rabbit antisera raised against 5 alpha-androstane-3 alpha, 11 alpha, 17 beta-triol 17-glucuronide 11-glutaryl bovine serum albumin and a heterologous tracer of androstanediol-17G conjugated with horseradish peroxidase at the glucuronic acid group were used. The EIA showed excellent specificity: there were no remarkable cross-reactivities with related androgens. The assay range for urine samples was 0.3-30 ng/ml. Recoveries of standards added to samples were 100-108%. Intra-assay and inter-assay coefficients of variation were 2.9-4.4% and 5.7-7.9%, respectively. The EIA was applied to urine samples of 407 males and 322 females to determine developmental patterns and normal ranges of androstanediol-17G excretions in 11 age groups (0 y, 1 y, 2-3 y, 4-5 y, 6-7 y, 8-9 y, 10-11 y, 12-13 y, 14-15 y, 16-17 y, and over 18 y). Urinary androstanediol-17G/creatinine (androstanediol-17G/Cre) ratios in both sexes were high in infancy, tended to decrease during childhood, and began to increase near adolescence. While androstanediol-17G/Cre ratio in girls increased at 8-9 y and reached a plateau during adolescence, that in boys increased at 10-11 y and continued to increase throughout adolescence. Androstanediol-17G/Cre ratios in girls were higher than those in boys at 6-7 y (P < 0.05) and at 8-9 y (P < 0.01). Androstanediol-17G/Cre ratios in boys were higher than those in girls at 12-13 y and at older ages (P < 0.01). These developmental patterns are parallel to age-related changes in androgenicity and serum androstanediol-17G, suggesting that urinary androstanediol-17G/Cre ratio could be a good marker for androgenicity in childhood.
authors.affiliation: Department of Human Ontogeny and Childhood Development (Pediatrics), Tokyo Medical and Dental University, Yushima 1-5-45, Bunkyoku, Tokyo 113-8519, Japan. tonishi@tmd.ac.jp
authors.forename: Toshikazu
authors.initials: T
authors.lastname: Onishi
authors.forename: Hajime
authors.initials: H
authors.lastname: Takei
authors.forename: Akira
authors.initials: A
authors.lastname: Kambegawa
authors.forename: Sumitaka
authors.initials: S
authors.lastname: Saisho
authors.forename: Kenichi
authors.initials: K
authors.lastname: Kashimada
authors.forename: Satomi
authors.initials: S
authors.lastname: Koyama
authors.forename: Shuki
authors.initials: S
authors.lastname: Mizutani
authors.forename: Pemmaraju N
authors.initials: PN
authors.lastname: Rao
mesh_terms: D000293:Adolescent; D000328:Adult; D015113:Androstane-3,17-diol / Q000031:analogs & derivatives* / Q000652:urine*; D002648:Child; D002675:Child, Preschool; D005260:Female; D006735:Horseradish Peroxidase; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D011786:Quality Control; D012016:Reference Values; D012680:Sensitivity and Specificity; D012727:Sex Characteristics
chemical_list: D015113:Androstane-3,17-diol; C028895:androstane-3,17-diol glucuronide; D006735:Horseradish Peroxidase