Developmental study for diagnostic techniques of congenital hepatobiliary disorders by fetal bile acids

点击翻译按钮获取中文摘要

To develop a diagnostic technique for congenital hepatobiliary disorders, qualitative and quantitative analysis of the conjugated 1beta- and 6alpha-hydroxy bile acids including common bile acids has been performed by high-performance liquid chromatography with chemiluminescence detection. Analysis of urine samples from newborns, normal women and women with pregnancy showed a large defderence in bile acid composition and conjugation mode, suggesting the distinctiveness of the bile acid metabolism during fetal and neonetal periods.Anti-tauro 1beta-hydroxycholic acid antisera have been prepared by immunizing rabbits with bovine serum albumin conjugates, in which the glycine and beta-alanine derivatives of 1 -hydroxycholic acid were coupled at the C-24 position. The antisera had high affinity and specificity for conjugated 1beta-hydroxycholic acid ; cross-reactivities for glyco 1beta-hydroxycholic acid were 100% and those for glycine and taurine conjugates ranged from 43.1 to 0.23%. Concentrations of conjugated 1beta-hydroxycholic acid were determined by radioimmunoassay with the more specific antiserum to be significantly large amounts in newborns at 0 - 20 days after birth, ranging from 0.29 to 18.51 mug/ml, when compared to normal women (<0.12 mug/ml) and women with late pregnancy (<1.13 mug/ml).To facilitate mass screening for congenenital biliary atresia, radioimmunoassay of conjugated 1 -hydroxycholic acid in dried blood spotted on filter paper was developed. The assay method was sufficiently sensitive and specific to determine the bile acid in one disc (3mm i. d., 4mul of whole blood). Preliminary tests suggested that the efficiency of this screening is acceptable, in which the levels of the patients (2.35 ng/disc) were significantly higher values than those of normal newborns (0.97 ng/disc).