Preliminary experiences with a bioassay for adrenocorticotrophin (ACTH) in unextracted human plasma using dispersed guinea-pig adrenal cells.
A simple, accurate and precise dispersed cell bioassay suitable for the assay of ACTH levels greater than 50 ng/1 in unextracted human plasma is described. Isolated adrenal cells were prepared by tryptic digestion of the guinea-pig adrenal gland and cortisol production by these cells was controlled specifically by ACTH. The cortisol response to 2 ng/1 of human pituitary ACTH (1-39) was always significantly (P less than 0.05, n = 17) different from the control. ACTH concentrations between 1 and 20 ng/1 gave a 14 +/- 3 fold (n = 17) increment in cortisol response. Dilutions of test plasmas were parallel with standard ACTH (1-39) when the plasma concentrations were maintained throughout at 4% (1 in 25 dilution) by the addition of dexamethasone-suppressed plasma ('ACTH free'). Using a 25 fold plasma dilution, the limit of detection of ACTH in unextracted plasma was 50 ng/1. Recovery of ACTH (1-39) spiked into dexamethasone suppressed plasma at concentrations of 250 and 100 ng/1 was 97 +/- 10% (n = 17) and 114 +/- 15% (n = 17) respectively. The within assay coefficient of variation (CV) of both quality control and patients' samples never exceeded 9% while between assay variation was less than or equal to 13%. The sample throughput was 30 plasma samples/week/technician.
D000224:Addison Disease / Q000097:blood; D000311:Adrenal Glands / Q000166:cytology; D000324:Adrenocorticotropic Hormone / Q000097:blood*; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D003480:Cushing Syndrome / Q000097:blood; D006168:Guinea Pigs; D006801:Humans; D009347:Nelson Syndrome / Q000097:blood
D000324:Adrenocorticotropic Hormone