Abstract
This study examined longitudinal, age-related and intra-individual variation in Anti-Müllerian Hormone (AMH) in regular menstruating women and correlated the hormonal levels to the antral follicle count (AFC). The impact of variations on an algorithm for calculation of follitropin-dose for ovarian stimulation were also tested. The study was carried out at a fertility clinic of a tertiary university hospital and had a prospective trial design. Twenty-six healthy women not receiving infertility treatment aged 22 to 50 years participated. Blood sampling for hormonal analysis was done every fifth day throughout three consecutive menstrual cycles, AFC was determined with 3-dimentional ultrasound and AMH measured by different assays from Beckman Coulter, Roche and Ansh Labs. Outcome measures were maximum and minimum difference in absolute and relative terms for each study subject during the test-period, coefficient of variation (Cv) for AMH for each cycle and cycle-day and correlation between AMH and AFC. The impact from variable AMH levels on an algorithm calculating follitrophin-delta dose in ovarian stimulation was explored. A significant longitudinal age-independent variation in AMH-levels and coefficient of variation in cycles and cycle days was found. A strong correlation between AMH-levels and AFC was confirmed and a case of significant divergence between assays was seen. Variations in AMH had a significant impact on an algorithm calculated dosage of gonadotrophins in ovarian stimulation. The finding of a substantial longitudinal variation in AMH question one recording being sufficient in quantifying gonadotrophins for ovarian stimulation, decision making and prognostication related to infertility treatment and counseling. Occasionally, commercial assays may fail to recognize specific AMH cleavage-products.
Highlights
The serum concentration of Anti-Müllerian Hormone (AMH) has gained widespread clinical use as a surrogate marker for ovarian reserve
The major finding of this study is a substantial overall intraindividual variation in AMH levels which to the best of our knowledge has not previously been reported with sample collection at 5 days intervals during three consecutive menstrual cycles taking only normally menstruating woman into account
In contrast to markers like FSH, no recommendations regarding a particular cycle day for optimal AMH assessment are currently recommended in the clinical context, distinct AMH cut off concentrations are often utilized to discriminate between normal and high or low values
Summary
The serum concentration of Anti-Müllerian Hormone (AMH) has gained widespread clinical use as a surrogate marker for ovarian reserve. The inter-assay coefficient of variation (%) was determined by measuring daily replicates of controls in two levels in 36 consecutive workdays, with the result of 1.9% at both levels (6.3 and 31.2 pmol/l) These two assays use 2 identical monoclonal antibodies directed against epitope regions located within the mature and pro-regions of the AMH molecule [21, 22]. One participant (#6) had low mean values of AMH including several measures below detection limit and high Cv-values This Participant was included in calculations but to investigate whether the inclusion of this participant changed the results sensitivity analyses excluding this were performed. All statistics were performed using R version 3.3.3 (R Foundation of Statistical Computing, Vienna, Austria)
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