The original Beckman Coulter Generation II assay significantly underestimates AMH levels compared with the revised protocol

Abstract

Purpose Anti-Müllerian hormone (AMH) is used as a marker for ovarian reserve. Since 2011, the standard test for AMH has been the Beckman Coulter Generation (Gen) II assay. However, in July 2013, the protocol was revised due to falsely low readings. The aim of this study was to compare AMH levels measured with the original and revised Gen II assay and to establish a fertile female reference range for the revised protocol. Methods Serum AMH levels were measured for 492 natural conception first trimester pregnant women using the original and revised Gen II assay. Results The original protocol significantly underestimated AMH levels compared with the revised protocol (p<0.001), the median being 8.4 and 14.2 pmol/L, respectively. In all samples with detectable AMH levels, the revised protocol yielded a higher concentration compared with the original protocol, the magnitude shift ranging from 3.4 to 283.3 % (median 68.0 %). AMH levels measured with the revised protocol were collated to generate an age-specific reference range, with median levels peaking at 27 years then declining with advancing age. The median AMH concentration for ages 20-24 was 17.3 pmol/L, ages 25-29 was 20.5 pmol/L, ages 30-34 was 17.8 pmol/L, ages 35-39 was 10.8 pmol/L, and ages 40-44 was 6.1 pmol/L. Conclusions Our study demonstrated that the original Gen II assay significantly underestimated AMH levels, suggesting caution is required when interpreting literature and testing results achieved with this assay. We also established the revised Gen II assay reference range for AMH in women with unassisted proven fertility.

Simplified 4‐item criteria for polycystic ovary syndrome: A bridge too far?

“…In July 2013, the Gen II assay protocol was revised to address concerns that results generated using an undiluted sample may potentially be falsely suppressed; fresh or freshly frozen specimens being particularly susceptible to complement interference. 26 Nevertheless, a recent study has demonstrated that AMH assay methods, manufactured by Beckman Coulter, Roche and Ansh Labs are well correlated. 27 A healthy cohort recruited from an annual heath screen enabled us to establish thresholds for testosterone, AFC and OV.…”

Paediatric Anti‐Müllerian Hormone measurement: Male and female reference intervals established using the automated Beckman Coulter Access AMH assay

“…18 In 2010, Beckman Coulter combined these technologies to produce a second-generation (Gen II) AMH ELISA. 19 This assay was subsequently modified to minimize interference from complement [20][21][22] and has recently been adapted for the Beckman Access automated platform.…”

Effect of Female Genital Tubercular Involvement on Anti-Mullerian Hormone Secretion in Latent Genital Tuberculosis Affected Women

“…Serial incubation & washing with anti-AMH detection antibody, enzyme and substrate enables to determine the enzymatic turnover of the substrate that is directly proportional to the concentration of AMH in the serum. According to our reference interval, median AMH concentration for ages 20-24 was 2.42 ng/ml, ages 25-29 was 2.87 ng/ml, ages 30-34 was 2.49 ng/ml, ages 35-39 was 1.51 ng/ml [12].…”

“…On the same day (2 nd day) venous blood samples were collected from those subjects for the measurement of AMH level. Serum and plasma samples were centrifuged at 1700g within 2 hours from the collection time and stored at -80 º C until assayed [12].…”