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Validation, normative values and clinical applicability of an enzyme immunoassay for the determination of serum anti-Müllerian hormone

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Author(s):
Renata dos Santos Batista Reis Woloszynek
Total Authors: 1
Document type: Master's Dissertation
Press: São Paulo.
Institution: Universidade de São Paulo (USP). Faculdade de Medicina (FM/SBD)
Defense date:
Examining board members:
Vinicius Nahime de Brito; Cristiane Kochi
Advisor: Vinicius Nahime de Brito
Abstract

The anti-Müllerian hormone (AMH) is a marker of ovarian reserve and testicular function. The clinical application of this hormone requires proper standardization of reference values according to the immunoassay used. The aims of this study were: to validate the AMH Gen II immunoassay (Beckman Coulter Company, TX, USA), to establish reference AMH values in healthy men and women, the influence of hormonal contraceptive use, smoking and body mass index (BMI) on the values of AMH and to check serum concentrations of this hormone in patients with Turner syndrome (TS), polycystic ovary syndrome (PCOS) and in boys with cryptorchidism underwent stimulation test rhCG (recombinant human chorionic gonadotropin).The validation of the AMH Gen II assay was performed using simplified protocol following the recommendations of the Clinical and Laboratory Standards Institute. To establish reference AMH values, 133 healthy women and 135 men were prospectively selected. In addition, 66 patients with TS, 29 with PCOS and 31 with cryptorchidism were studied. The analytical and functional sensitivity of the AMH Gen II assay was 0.02 and 0.2 ng / mL, respectively. Intra and inter-assay coefficients of variation in different concentrations ranged between 5.2-9.0% and 4.6-7.8%, respectively. The linearity, parallelism and stability studies showed recovery % between 80 and 120%. The AMH Gen II assay strongly correlated with the previously used Immunotech assay (r = 0.9, p < 0.001). In females, serum AMH showed a progressive decline with increasing age (r = -0.4, p < 0.001). On the other hand, AMH values did not differ between users and nonusers of hormonal contraceptives, smokers and nonsmokers, obese and non-obese. In males, age did not influence the levels of serum AMH, however, AMH values were significantly reduced with increasing BMI (r = -0.3, p = 0.008), but, these values were within the range observed in the group of subjects with body weight. All TS patients showed undetectable AMH levels. In contrast, in PCOS AMH values were significantly higher than in women controls, but with overlap of 76% between the two groups. In boys with cryptorchidism, the concordance between baseline AMH and testosterone after stimulation with rhCG was 74%, there was a significant positive correlation between baseline AMH values and testosterone values after stimulation (r = 0.5, p < 0.001). In conclusion, AMH Gen II assay is reliable for determining the serum AMH. Reference AMH values showed a wide range in both sexes. Reduced values of AMH in obese men were within the range of normal individuals. In ST, the potential use of AMH to assess ovarian reserve requires longitudinal studies. The use of AMH as a diagnostic criterion of SOP must be based on the association with the criteria already established. In boys with cryptorchidism, a normal AMH provides useful information on testicular function, but does not exclude the need for stimulation test rhCG, patients with discordant results require follow-up (AU)

FAPESP's process: 12/20525-8 - Validation, normative date and clinical utility of and enzymeimmunoassay for serum measurement of anti-Müllerian hormone
Grantee:Renata dos Santos Batista Reis Woloszynek
Support Opportunities: Scholarships in Brazil - Master