Detection of serum antimüllerian hormone in women approaching menopause using sensitive antimüllerian hormone enzyme-linked immunosorbent assays.
OBJECTIVE
Current antimüllerian hormone (AMH) immunoassays are insufficiently sensitive to detect circulating AMH levels in ovulatory women approaching menopause. The aim of this study was to detect serum AMH levels across the menstrual cycle with age, using two new AMH enzyme-linked immunosorbent assay (ELISA) kits with increased sensitivity and differing specificity.


METHODS
Serum AMH levels were determined every 2 to 3 days across the interovulatory interval of menstrual cycles among women of early-mid reproductive age (18-35 y; n = 10) and late reproductive age (45-55 y; n = 17). Two highly sensitive AMH ELISAs (designated 24/32 and 24/37) with differing sensitivities were developed and applied to sera using a recombinant human pro-mature AMH preparation as reference. A third AMH ELISA (Gen II AMH ELISA kit; Beckman Coulter, Brea, CA) used was directed on mature-pro regions of AMH.


RESULTS
AMH levels in all cycles were detectable with the 24/32 and 24/37 AMH ELISAs. AMH levels across the menstrual cycle were highly correlated (r = 0.98) between the 24/32 and 24/37 AMH ELISAs and the Gen II AMH ELISA (r = 0.94), but with large intracycle variations observed in older women. In late reproductive age, more than 95% of AMH values were detectable with the 24/32 and 24/37 AMH ELISAs, whereas only 36% of AMH values were detectable with the Gen II AMH ELISA. AMH levels were detected in cycles with lower antral follicle count and at a later age using the 24/32 and 24/37 AMH ELISAs compared with the Gen II AMH ELISA. AMH level correlated with antral follicle count in younger women, but not in older women.


CONCLUSIONS
The new 24/32 and 24/37 AMH ELISAs have the sensitivity to monitor ovarian follicle profiles in late reproductive age.
From the 1Monash Institute of Medical Research-Prince Henry's Institute of Medical Research, Clayton, Victoria, Australia; 2Department of Obstetrics and Gynecology, Monash University, Clayton, Victoria, Australia; 3Ansh Labs, Webster, TX; 4Department of Obstetrics, Gynecology, and Reproductive Sciences, College of Medicine, University of Saskatchewan, Saskatchewan, Canada; and 5Department of Internal Medicine, Erasmus MC, Rotterdam, the Netherlands.
David M
DM
Robertson
Ajay
A
Kumar
Bhanu
B
Kalra
Shivani
S
Shah
Enid
E
Pruysers
Heidi
H
Vanden Brink
Donna
D
Chizen
Jenny A
JA
Visser
Axel P
AP
Themmen
Angela
A
Baerwald
D000293:Adolescent; D000328:Adult; D000367:Age Factors; D054304:Anti-Mullerian Hormone / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D008593:Menopause / Q000097:blood*; D008597:Menstrual Cycle / Q000097:blood*; D008875:Middle Aged; D006080:Ovarian Follicle; D010053:Ovary / Q000502:physiology; D012680:Sensitivity and Specificity; D055815:Young Adult
D054304:Anti-Mullerian Hormone