Modification of the Beckman Coulter second generation enzyme linked immunosorbent assay protocol improves the reliability of serum antimüllerian hormone measurement

The modified second-generation enzyme-linked immunosorbent assay protocol eliminates potential intrinsic and extrinsic interference and gives more reliable results on sample dilution and storage compared with the original second-generation protocol.

Laurentiu Craciunas, Dr.-Med., Stephen A. Roberts, Ph.D., Allen P. Yates, Ph.D., Alexander Smith, Ph.D., Cheryl Fitzgerald, M.D., Philip W. Pemberton, M.Sc.

Volume 103, Issue 2, Pages 553-559


To determine whether the modified Beckman-Coulter 2nd-generation (Gen II) antimüllerian hormone (AMH) assay (Gen IIm) provides more consistent results following storage at room temperature and on dilution than the original Gen II assay, to compare AMH results from the modified assay with those obtained from the original assay, and to assess the relationship between new AMH values and the antral follicle count (AFC).


Hospital fertility clinic.

A total of 678 consecutive women (21–46 years old) investigated for subfertility.


Main Outcome Measure(s):
AMH was measured by means of the Gen IIm assay protocol in women with known AFC. AMH values were obtained on a subset of serum samples by means of both original and modified assays.

Specimens analyzed by Gen IIm exhibited a proportional AMH response on dilution, and AMH values decreased by an average of 12.1% after 7 days at room temperature, in contrast to the steady increase seen with the use of the original Gen II assay. Gen IIm assay values were, on average, 51.4% higher than Gen II values. Population analysis suggested a conversion factor of 1.35 (95% CI 1.23–1.47) between the Gen IIm and historical data obtained for the Diagnostic Systems Laboratories AMH assay. The relationship between the Gen IIm AMH measurement and AFC was adequately represented by a linear function.

The Gen IIm assay gave more reliable AMH results on sample dilution and storage than the original Gen II protocol. Findings obtained with the use of the original Gen II ELISA method should be treated with caution.

  • Shvetha Zarek

    Although AMH is widely used in clinical practice as a marker of ovarian reserve, issues related to the validity of the assay continue to hamper reliable intepretation of values and the ability to compare studies. The authors undertake the important task of evaluating if the modification of the Beckman Coulter assay that was implemented in August 2013 has improved reliability. The authors evaluated follicular phase AMH samples in 678 women ages, 21-46 years, and demonstrated that modified Gen IIm assay gave values that were 51.4% higher than the original Gen II assay. Overall, samples at room temperature demonstrated improved stability with the modifed assay. The authors commendably also evaluated the relationship between antral follicle count and AMH values with the modified assay. Supplemental Figure 1 provides vital information that is perhaps more useful than the data provided by the age and AMH comparison in Figure 2. Although there is an overall linear relationship between AFC and AMH, it seems that there is a wide range of AMH values for the same AFC value and some quite low AMH values with normal AFCs. Can the authors kindly further comment on their findings in supplemental Figure 1?

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