Validation of an immunoassay for anti-Mullerian hormone measurements and reference intervals in healthy Brazilian subjects

Abstract

Background Anti-Mullerian hormone is marker of ovarian and testicular reserve. The clinical use of this hormone requires proper standardization of reference intervals. The aims of this study were to validate the Anti-Mullerian hormone Gen II immunoassay, to establish Anti-Mullerian hormone reference intervals in healthy subjects, and to evaluate the influence of hormonal contraceptives, smoking, and body mass index on Anti-Mullerian hormone. Methods The validation of the Anti-Mullerian hormone Gen II assay (Beckman Coulter Company, TX, USA) was performed using a simplified protocol recommended by Clinical Laboratory Standard Institute. One-hundred and thirty-three healthy females and 120 males were prospectively selected for this study. Results The analytical and functional sensitivities of the Anti-Mullerian hormone Gen II immunoassay were 0.02 and 0.2ng/mL, respectively. Intra-assay coefficients ranged from 5.2 to 9.0%, whereas inter-assay precision ranged from 4.6 to 7.8% at different concentrations. In females, Anti-Mullerian hormone showed progressive decline with increasing age (r=-0.4, p<0.001), whereas in males, age showed no influence on Anti-Mullerian hormone concentrations. In females, Anti-Mullerian hormone concentrations did not differ between users and non-users of hormonal contraceptives, smokers, and non-smokers and obese and lean individuals. However, there was a negative and significant correlation between Anti-Mullerian hormone and body mass index in males (r=-0.3, p=0.008). Conclusions Anti-Mullerian hormone Gen II assay was reliable for determining serum Anti-Mullerian hormone concentrations. Anti-Mullerian hormone concentrations declined with aging and presented a wide inter-individual variability. The lack of influence of hormonal contraceptives, smoking, and obesity on Anti-Mullerian hormone in both sexes allowed us to refine the normative concentrations for the Brazilian population.