AMH Assay Research Articles

Hormone anti-müllérienne (AMH) en pédiatrie: établissement de valeurs de référence avec le dosage de l'AMH Fujirebio® Lumipulse G et comparaison avec le dosage Roche® Elecsys sur les mêmes échantillons pédiatriques

We performed a method comparison between the Fujirebio® Lumipulse G AMH assay and the Roche® Elecsys AMH assay using the same pediatric samples. We described full pediatric gender and age-specific reference ranges for AMH using the Fujirebio® AMH assay on the Lumipulse G 600 II. The study was performed on 281 plasma samples collected in tubes with lithium heparin. The samples were from patients (135 males and 146 females) aged from 3 days to 22 years collected at the University Hospital Center of Tours. The Fujirebio® Lumipulse method showed excellent correlation with Roche® Elecsys but had a significant proportional positive bias. The data were used to propose pediatric reference values adapted to the Fujirebio® method. Our study described full pediatric gender and age-related reference ranges for AMH using the Fujirebio® AMH assay on the Lumipulse G600II. The delineation between normal male and female AMH concentrations make them valuable clinical tools for the monitoring of pediatric sexual and reproductive development from early childhood through the pubertal transition into adulthood.

O-067 Does the measurement of different AMH isoforms in poor responder patients improve the prediction of ovarian response?

Abstract Study question Does the measurement of AMH isoforms in low ovarian reserve patients improve the prediction of the number of oocytes after ovarian stimulation (OS)? Summary answer AMH assays measuring a broader range of AMH isoforms combined with antral follicle count (AFC) improve the prediction of oocytes collected after OS. What is known already Granulosa cells secrete AMH as a non-active prohormone (proAMH). After cleavage, it forms a complex (AMHN,C) which activates the receptor (AMHR2). Circulating AMH is a mixture of isoforms (proAMH, AMHN,C and other sub-fragments) after proteolysis, targeted by the monoclonal antibodies included in the AMH assays commonly used in clinical practice, potentially affecting the quantification of the hormone. Novel AMH assays include antibodies directed towards a broader range of the molecule. They may provide more accurate and reliable results, especially important in poor responders, as results may involve clinical decisions on whether or not to proceed with OS. Study design, size, duration Prospective observational study measuring AMH isoforms using five different AMH assays, including 72 women with low ovarian reserve performing OS for IVF/ICSI at a tertiary referral fertility center, from February 2019 to December 2021. Low ovarian reserve was defined as AMH serum levels <1.1ng/ml (Elecsys, Roche), following Bologna criteria. All patients performed OS for IVF/ICSI with antagonist protocol. Participants/materials, setting, methods On day 2/3 of cycle, prior to initiating the OS, AFC, serum FSH, LH, estradiol, progesterone and AMH levels were measured using Elecsys assay (Roche). Extra serum samples were frozen at -20C for subsequent analysis, using four different AMH assays (AnshLabs, Texas): AL-196 (PCOCheck ELISA), AL-124 (PicoAMH ELISA), AL-105 (US-AMH ELISA) and AL-133 (Total Mature-AMH ELISA). Ethical approval was obtained (Research Ethics Committee-REFA033c) and informed consent was signed by all participants. Main results and the role of chance Patient characteristics were [Median(IQR)]: age:39(36-42)years, BMI:28.51(26.1-30.1) Kg/m2, AFC:5.5(4-7), FSH:8.23(6.4-12.5) mIU/mL, LH 6.54(4.51-8.91) mIU/mL, Estradiol: 41.18(28.47-56.44)pg/mL, AMH-Elecsys: 0.64(0.29-0.81)ng/mL, AL-196:0.64(0.39-0.99)ng/mL, AL-124:0.79(0.49-1.24ng/mL, AL-105: 0.89(0.57-1.39)ng/mL, AL-133:1.06(0.63-1.59)ng/mL. Stimulation outcomes were: follicle number on the day of trigger (Fdot):5(3-7), cumulus-oocyte-complexes (COC’s):3(2-4.5) and metaphase II oocytes (MII):3(1-4). Spearman correlation was performed between AMH assays, AFC and OS outcomes. Elecsys revealed a good correlation with the other AMH assays (rs = 0.60-0.63,p<0.001), however, it was higher among ELISA assays (rs = 0.954-0.993,p<0.001). All AMH assays showed a significant positive correlation with AFC, Fdot, COC’s and MII. However, AL-196 showed the highest correlation (rs = 0.524, rs = 0.615, rs = 0.594, rs = 0.595, respectively;p<0.001). Different models combining one AMH assay plus AFC were created to predict COC’s and MII: AFC+AL-196 revealed the best prediction (Adjusted R2=0.474[p < 0.001] and 0.485[p < 0.001], respectively) when compared to the same AMH assay alone. AUROC analysis was performed to investigate which model predicted better < =3COC’s after OS. Although all models demonstrated fair values, AUROC was better for AFC+AL-196 model (AUROC:0.796), yet no significant differences were seen when compared to AUROC for AFC+AMH-Elecsys (p = 0.39). AFC and AMH were good predictors for COC’s collected [RR(95%CI):1.49(1.11-2.04),p<0.01; RR(95%CI):1.26(1.13-1.49),p<0.001, respectively). AFC+AMH-Elecsys predicted COC’s better than AFC alone (R2=0.47 vs 0.29, respectively;Performance-Score:42.56% vs 13.21%, respectively). AFC+AL-196 had the highest performance (R2=0.599;Performance-Score:90.12%). Limitations, reasons for caution Although clinical assessment was performed in the same centre following the same methodology, inter-observer variability for AFC is a limitation. Besides, fresh AMH serum samples were analysed using Elecsys assay, whereas frozen serum samples were shipped to Ansh Lab (Texas) for batched analysis. Wider implications of the findings For patients with serum AMH<1.1ng/mL, a wide range of oocytes might be expected after OS. Novel AMH assays, targeting a wider broad of the molecule, together with AFC, can be used for these patients to obtain a better prediction for clinical outcomes and anticipate very poor response. Trial registration number Not applicable

P-657 Elecys anti-Müllerian hormone thresholds for defining ovarian response categories; comparison of published and novel values in 7,988 treatment naïve women

Abstract Study question Test the accuracy of published and novel AMH thresholds in predicting (low, sub-optimal, optimal and high) ovarian response categories in first treatment cycles. Summary answer Published AMH thresholds for poor and excessive ovarian response perform adequately, however, prediction of suboptimal or optimal oocyte yields is difficult reflecting their limited range. What is known already AMH is an established predictor of ovarian response, and can identify women at risk of poor (<4 oocytes) or excessive (>15 oocytes) response. With the development of suboptimal (4 to 9 oocytes) and optimal (10 to 14 oocytes) response categories, a series of AMH ranges for the Roche Elcesys AMH assay has been proposed to predict response for all four categories (AMH <6.4pmol/l, 4.9–11.3 pmol/l, 11.3–20.9 pmol/l, and >14.2 pmol/l). To date there has been no internal or external validation of these single centre derived thresholds, and it is unknown whether they are generalisable to other clinic populations. Study design, size, duration Observational cohort study from 8 UK clinics, incorporating 7,998 women undertaking their first ovarian stimulation cycle for assisted conception or fertility preservation between 2016 and 2022, with a pre-treatment AMH measured using the Roche Elecsys AMH assay. Both GnRH agonist and antagonist cycles were included, with triggering by either recombinant hCG or GnRH agonist once 3 follicles were ³18mm in size with oocyte retrieval performed 36h later. Participants/materials, setting, methods To give equal opportunity to the published and novel thresholds, patients were divided into training (5,330) and testing (2,668) sets. Novel cut-off points for a low, sub-optimal, optimal and high response were derived in the training set. The performance of these novel thresholds and those of published values were then assessed and compared in the testing set by their respective AUC, and precision as defined as the fraction of correct predictions for a certain class. Main results and the role of chance Baseline patient characteristics (mean, SD) were 35.1±4.7 years; BMI 25.5±4.4 kg/m2, with a median AMH 15.2 pmol/l (IQR 8, 26) and a median of 11 (IQR 6,16) oocytes retrieved. These baseline characteristics and outcomes were similar after splitting in the training and testing datasets. Baseline AMH and oocytes collected were moderately correlated (r = 0.54, p < 0.001). 465 (5.8%) cycles were cancelled due to not meeting trigger criteria and classed as poor/low responders. The novel thresholds for low oocyte yield (≤10.05 pmol/l) provided similar performance (AUC 0.70 95%CI 0.66-0.74, sensitivity 0.69, specificity 0.74) to the published thresholds (AUC 0.67, 95%CI 0.62-0.72) in the testing population. Similarly, the novel high (15.9pmol/l) threshold (AUC 0.78, 95%CI 0.75-0.81, sensitivity 0.78, specificity 0.64) exhibited similar performance to the published threshold AUC 0.77, 95%CI 0.74,0.81) in the testing population. Deriving novel thresholds for prediction of 4 categories of response (6.7, 12.9, 18.9pmol) was associated with slightly better precision for low, suboptimal and high categories (p < 0.001), with the fraction of correct predictions for a certain category of response for the novel cut-points; 69.0 % low, 11.7% suboptimal, 14.1% optimal and 78.3% high, as compared to 52.0%, 27.8%, 12.9% and 83.8% respectively for the published values. Limitations, reasons for caution While designing the ovarian stimulation strategies clinicians were aware of the baseline AMH and targeted a perceived optimal response for cumulative live birth (∼15 oocytes), which may have contributed to the performance of both the established and novel AMH thresholds. Wider implications of the findings In a large heterogenous population of women undergoing their first treatment cycle, we confirm the appropriateness of the previously published Elecsys AMH values for prediction of low (<6.4pmol/l) and high (>14.2pmol/l) oocyte yields. Trial registration number N/A

USE OF ELECTROCHEMILUMINESCENCE METHODS TO EVALUATE THE RELATIONSHIP BETWEEN OVARIAN RESPONSE AND ANTI-MULLERIAN HORMONE (AMH) LEVELS WHEN THEY ARE LOWER THAN 1 NG/ML

The evolution of biochemical methods to measure AMH allowed to obtain increasingly precise and reliable values of this marker of ovarian reserve. The Elecsys® AMH assay is an ELISA-type immunoassay based on electrochemiluminescence (EQL) technology and represents one of the most robust diagnostic methods for measuring this hormone. Elecsys® AMH assay could allow the analysis of the relationship between the number of oocytes obtained in a follicular puncture after controlled ovarian stimulation (COS) with AMH levels, when they are lower than 1 ng/ml, a value below which the ovarian response could not be predicted with sufficient confidence. We aimed to establish the diagnostic sensitivity of the Elecsys°AMH method when the hormone level is less than 1 ng/ml in relation to the number of oocytes recovered in follicular punctures. Specifically, determine if there are differences in the number of oocytes obtained depending on the concentration of AMH in the sample: less than 0.49 ng/ml and between 0.50 – 0.99 ng/ml

Evaluation of at-home serum anti-Müllerian hormone testing: a head-to-head comparison study

BackgroundFor optimal fertility testing, serum anti-Müllerian hormone levels are used in combination with other testing to provide reliable ovarian reserve evaluations. The use of the ADx 100 card is widely commercially available for at-home reproductive hormone testing, but data demonstrating that its results are reproducible outside of a clinical setting are limited, as well as comparisons of its performance with other newer blood collection techniques. This study aimed to evaluate the concordance of serum AMH levels found via standard venipuncture and self-administered blood collection using the TAP II device (TAP) and ADx card in women of reproductive age.MethodsThis was a prospective, head-to-head-to-head within-person crossover comparison trial that included 41 women of reproductive age (20–39 years). It was hypothesized that the TAP device would be superior to the ADx card both in terms of agreement with venipuncture reference standard and patient experience. Each subject had their blood drawn using the three modalities (TAP, ADx, and venipuncture). We evaluated the concordance of AMH assays from samples obtained via the TAP device and ADx card with the gold standard being venipuncture. Two-sided 95% CIs were generated for each method to compare relative performance across all three modes. Patient preference for the TAP device versus the ADx card was based on self-reported pain and Net Promoter Score (NPS).ResultsThe TAP device was superior to the ADx card on all outcome measures. TAP R-squared with venipuncture was 0.99 (95% CI 0.99, > 0.99), significantly higher than the ADx card, which had an R-squared of 0.87 (95% CI 0.80, 0.94) under most favorable treatment. TAP sensitivity and specificity were both 100% (no clinical disagreement with venipuncture), versus 100 and 88%, respectively, for the ADx card. Average pain reported by users of the TAP device was significantly lower than the ADx card (0.75 versus 2.73, p < 0.01) and the NPS was significantly higher than the ADx card (+ 72 versus − 48, p < 0.01).ConclusionsThe TAP was non-inferior to venipuncture and superior to the ADx card with respect to correlation and false positives. Moreover, the TAP was superior to both alternatives on patient experience.Trial registrationNCT04784325 (Mar 5, 2021).

Comparison of three different AMH assays with AMH levels and follicle count in women with polycystic ovary syndrome

Comparison of three different AMH assays with AMH levels and follicle count in women with polycystic ovary syndrome

Evaluating Mirna-222 Expression Level and Its Association with AMH for Early Diagnosis of Pcos-Like Symptoms in Epileptic Patients Plasma Treated with Sodium Valproate: A Case – Control Study

Introduction: Epileptic neurological disorder, which is controlled with medications such as sodium valproate (one of the treatment priorities for the patients with epilepsy). Polycystic ovary syndrome (PCOS) is one of complication of sodium valproate. Ovulation in PCOs Patients is disrupted, resulting in serious complications, including endometrial hyperplasia with typical and atypical forms, increased the risk of endometrial endometrial cancer, diabetes mellitus, and decreased fertility rates. This study, in accordance with bioinformatics studies and other studies based on the association of miRNA-222 with genes involved in PCOs and hyperandrogenism, was designed to evaluate the association of PCOs in the patients treated with sodium valproate by simultaneously comparing the AMH factor and miRNA-222 marker at specific time periods. Methods: In this case-control study, 33 women with epilepsy before and after use the drug were selected based on inclusion and exclusion criteria. After blood sampling, their plasma was isolated. According to the instructions of the Total RNA extracted kit , cDNA synthesized and miRNA-222 expression was evaluated by RT-qPCR technique and statistical analysis was performed by SPSS 23 software, T-test and ANOVA tests. Results: The results of statistical tests such as T-test and ANOVA test (done with SPSS software) which showed a significant difference (p&lt;0/01) between the mean expression of mir-222 and AMH in the patients before treatment compared to 3 month after treatment. The results of Pearson correlation test showed that the increase in AMH is directly related to the decrease in miRNA-222 expression (p&lt;0/01). Conclusion: According to the results of the present study, in order to better control the possible side effects of the drug and possibly timely drug change and early diagnosis of PCOs (PCOs‌-like symptoms), evaluation of miRNA-222 expression changes can be used at the same time with AMH assay.

Anti-müllerian hormone levels and antral follicle count in women with a BRCA1 or BRCA2 germline pathogenic variant: A retrospective cohort study

BackgroundSome studies suggested a decreased ovarian reserve among BRCA1/2 pathogenic variant carriers, with conflicting results. MethodsWe conducted a retrospective single-center observational study of ovarian reserve and spontaneous fertility comparing BRCA1/2 pathogenic variant carriers to controls (women who attended consultations to discuss fertility preservation before gonadotoxic treatment). Measures of associations between plasma AMH concentration, AFC and BRCA1/2 status were modelled by nonlinear generalized additive regression models and logistic regressions adjusted for age at plasma storage, oral contraceptive use, body mass index, cigarette smoking, and the AMH assay technique. ResultsThe whole population comprised 119 BRCA1/2 pathogenic variant carriers and 92 controls. A total of 110 women (42 carriers, among whom 30 were cancer-free, and 68 controls) underwent an ovarian reserve evaluation. Spontaneous fertility analysis included all women who previously attempted to become pregnant (134 women).We observed a tendency towards a premature decrease in ovarian reserve in BRCA1/2 pathogenic variant carriers, but no difference in mean AMH or AFC levels was found between BRCA1/2 pathogenic variant carriers and controls. An analysis of the extreme levels of AMH (≤5 pmol/l) and AFC (≤7 follicles) by logistic regression suggested a higher risk of low ovarian reserve among BRCA1/2 pathogenic variant carriers (adjusted odds ratio (OR) = 3.57, 95% CI = 1.00–12.8, p = 0.05; and adjusted OR = 4.99, 95% CI = 1.10–22.62, p = 0.04, respectively). DiscussionAttention should be paid to BRCA1/2 pathogenic variant carriers’ ovarian reserve, considering this potential risk of premature alteration.

MOTHER Study: A Multicenter Observational, Retrospective Study to Determine Coorelation Between Physical CHaracteristics and Ovarian REserve Markers in Sub-feRtile Women.

The physical characteristics which are known to affect the ovarian reserve are age, body mass index (BMI), occupational exposures, age at menarche and menstrual cycle length. A correlation between different physical characteristics and the ovarian reserve will help to identify areas which need to be tackled to increase the chances of fertility of women in India. In this retrospective, observational study, namely the MOTHER Study, data of women between 18 and 45years of age, attending the selected fertility centers across different states in India were taken for evaluation. Demographic information along with information on factors potentially related to fertility like age of menarche, menstrual cycle length and occupational factors were collected by review of medical records at screening visit. Most recent AMH assay and antral follicle count (AFC) where the subject has not taken any contraceptives 12months prior to the test were collected. Age of woman, years of marriage, years of infertility and smoking have shown effect on ovarian reserve testing like AMH and AFC. The other physical characteristics which were evaluated and considered to affect the ovarian reserve like body mass index BMI, occupational exposures, age at menarche and menstrual cycle length have not shown statistically significant correlation. Age of woman and years of infertility are inversely proportional to ovarian reserve markers, namely AMH and AFC. Addictions like smoking and alcohol affect ovarian reserve.

Androgens and Anti-Müllerian Hormone in Infertile Patients.

To investigate whether there is an association between androgens and ovarian reserve, expressed through anti-Mullerian hormone. This is a retrospective cross-sectional analysis of all consecutive women attending a tertiary fertility center, who presented with regular menstrual cycles. Patients had their AMH values measured with the same AMH assay (Immunotech (IOT) Beckmann Coulter assay), the same day in which androgens sampling was performed. Women with PCOS or other forms of androgen excess or untreated endocrine or metabolic disorders were excluded. A total of 942 women were included. Significant correlation was observed between total testosterone/free androgens index (FAI)/DHEAS and AMH (Spearman's r = 0.20/0.14/0.13, P value < 0.001, P value < 0.001, and P value < 0.001, respectively). After multiple linear regression analysis adjusting for confounders (age, BMI, cause of infertility, day of the menstrual cycle when the blood sample was performed), the regression slope in all participants for total testosterone predicting logAMH was 0.20 (P value < 0.001). Similarly, FAI was significantly associated with logAMH (regression coefficient = 0.04, P value = 0.04). In contrast, DHEAS was not significantly associated with logAMH. There was a significant, but weak relation between testosterone and AMH, while no significant association was observed between DHEAS and AMH. Future research is needed to elucidate whether testosterone supplementation may have any effect on ovarian function.

Differing performance of two assays for the measurement of anti-Mullerian hormone in premenopausal women: A cross-sectional study.

To compare the performance of two anti-Mullerian (AMH) assays over a range of concentrations, in samples collected from young women. A cross-sectional method-comparison study of 168 non-healthcare-seeking women. Included women were aged 18-39years, not recently pregnant, breast feeding or using systemic hormones. Serum AMH levels were analysed with the Beckman Coulter Access 2 assay from fresh samples and the Ansh picoAMH assay using samples stored at -80°C, in a parallel setting. Comparisons between the two assays were examined using Bland-Altman plots. Participants had a mean±SD age of 32.6±5.4years and body mass index of 28.1±7.9kg/m2 , and 60.1% were parous. Although the assay results were highly correlated (Spearman correlation .982, P<.001), the relationship between the assays was nonlinear. Serum AMH values below 4pmol/L were lower with the picoAMH assay compared with the Access AMH assay (mean difference in this range was -0.49pmol/L), but for samples with a mean value above 10pmol/L, the picoAMH assay consistently measured higher than the Access AMH assay (mean difference in this range was +8.2pmol/L). As AMH concentrations increased the absolute discrepancy between the assays also increased. This study demonstrates that despite the high correlation between two commercially available AMH assays, the assays performed in a discordant manner at high and low concentrations. Hence, the results of these assays are not interchangeable, highlighting the need to establish specific reference limits for individual assays to guide clinical decision-making and the challenge of establishing future universal cut-offs for the application of AMH levels in clinical practice.

Evaluation of ovarian reserve before and after chemotherapy

BackgroundProgress in oncology has improved patient survival. However, cancer chemotherapy can be gonadotoxic and affect their fertility. Recourse to fertility preservation before starting these treatments is therefore necessary in order to allow a better life quality after survival. The aim of this work was to study the impact of chemotherapy on ovarian reserve by AMH measurement. MethodsThis is a descriptive and longitudinal study from 2015 to 2018 carried out at Aziza Othmana hospital ART center in Tunis on patient aged less than 41 years who were candidates for fertility preservation. Patients included had AMH measurement prior to cancer treatment. We called them back to follow up the AMH level after chemotherapy. The AMH assay was performed by electrochemilumiescence technique. At the end, only 66 patients met the inclusion criteria. ResultsThe most frequent pathologies were Hodgkin's lymphoma and breast cancer. The mean age of patients was 26.7 ± 6.8. The most used chemotherapy protocols were BEACOPP, ABVD or the combination of both in lymphoma and FEC + TXT for breast cancer treatment. A significant difference between AMH before and after chemotherapy was found for BEACOPP and FEC + TXT protocols (p < 10 3). The patient’s age was correlated with the AMH decrease after chemotherapy (r = 0.577, p < 10 3). ConclusionOur results showed that the high risk gonadotoxicity protocols were BEACOPP for lymphoma treatment and FEC + TXT for breast cancer treatment. However, studies with a larger sample and more time extended monitoring are necessary for a better gonadotoxicity understanding of the cancer treatments available today.

AMH assessment five or more years after an initially low AMH level

PurposeAnti-Müllerian hormone (AMH) is a biomarker reflecting ovarian reserve. A low AMH level before 39 years may be associated with a risk of menopause between 40 and 45 years. The risk of onset of premature ovarian insufficiency (POI) is, however, poorly documented. The objective of the study was to determine the prevalence of POI 5–10 years after an AMH assay below 8 pmol/L. MethodsWe included women aged younger than 36 years who underwent a complete workup for infertility between January 2008 and December 2013 at the University Hospital and had an AMH level less than 8 pmol/L. In 2018, 47 of these women were assessed clinically, and 21 of them agreed to undergo an ovarian reserve test. ResultsThe prevalence of POI after at least 5 years was 8/47, or 17 % [8 %–31 %]. The median time to diagnosis was 5.1 years [2.9–7.3]. There was a significant difference at T0 in the regularity of cycles between the two groups (p = 0.024) and in their baseline serum AMH level: 6.0 pmol/L in the non-POI group vs 4.2 pmol/L in the POI group (p = 0.002). ConclusionSerum AMH < 8 pmol/L before the age of 36 years appears to be a risk factor for POI. These women require regular follow-up, especially if their cycles are irregular.

Anti-Müllerian Hormone and Ovarian Reserve: Update on Assessing Ovarian Function.

ContextAnti-müllerian hormone (AMH) is produced by granulosa cells of small, growing follicles in the ovary. Serum AMH levels strongly correlate with the number of growing follicles, and therefore AMH has received increasing attention as a marker for ovarian reserve. This review summarizes recent findings and limitations in the application of serum AMH in ovarian reserve assessment.Evidence AcquisitionA PubMed search was conducted to find recent literature on the measurements and use of serum AMH as a marker for ovarian reserve.Evidence SynthesisSerum AMH levels are measured to assess the “functional ovarian reserve,” a term that is preferred over “ovarian reserve,” since AMH levels reflect the pool of growing follicles that potentially can ovulate. Serum AMH levels are used in individualized follicle-stimulating hormone dosing protocols and may predict the risk of poor response or ovarian hyperstimulation syndrome but has limited value in predicting ongoing pregnancy. Serum AMH levels are studied to predict natural or disease-related age of menopause. Studies show that the age-dependent decline rates of AMH vary among women. The generalized implementation of serum AMH measurement has also led to an increase in diagnostic assays, including automated assays. However, direct comparison of results remains problematic.ConclusionSerum AMH remains the preferred ovarian reserve marker. However, the lack of an international standard for AMH limits comparison between AMH assays. Furthermore, little is known about endogenous and exogenous factors that influence serum AMH levels, which limits proper interpretation of AMH values in a clinical setting.

Prediction of ovarian response with an automated AMH assay (Elecsys®) in GnRH antagonist cycles

Prediction of ovarian response with an automated AMH assay (Elecsys®) in GnRH antagonist cycles

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

SummaryObjectiveAnti‐Müllerian Hormone (AMH) concentration is high at birth in males, demonstrating the presence of functional testicular tissue in the prepubertal period, and acting as a useful marker in the investigation of paediatric reproductive disorders. AMH also provides a tool in the investigation of female virilization, premature ovarian failure and polycystic ovarian syndrome in childhood. Robust, assay‐specific paediatric AMH reference intervals are therefore required for clinical interpretation of results. The aim of this study was to derive age‐specific AMH reference intervals for males and females aged 0‐18 years.Design and PatientsPlasma samples were obtained from patients at Royal Manchester Children's Hospital and analysed for AMH using the automated Beckman Coulter Access AMH Assay. Patients under investigation for paediatric reproductive or endocrine disorders were excluded from the study.MeasurementsSeven hundred and 2 patient plasma samples (465 male, 237 female) were subject to AMH measurement, and results were analysed in order to derive continuous and discrete reference intervals for the paediatric age range.ResultsClear discrimination between male and female AMH results was evident in the prepubertal age range, with some overlap between the genders following pubertal onset.ConclusionsWe have derived age‐related reference intervals for plasma AMH in the paediatric age range (0‐18 years) using the automated Beckman Coulter Access AMH assay which will aid in the investigation of paediatric endocrine disorders such as disorders of sexual development.

Multi-center clinical evaluation of the Access AMH assay to determine AMH levels in reproductive age women during normal menstrual cycles.

AMH is widely used for assessing ovarian reserve, and it is particularly convenient, because it is thought to have minimal variability throughout the menstrual cycle. However, studies assessing the stability of AMH over the menstrual cycle have been conflicting. The purpose of this study is to determine whether AMH levels vary across the normal menstrual cycle. A multi-center, prospective cohort study conducted at three US centers. Fifty females with regular menstrual cycles aged 18-45 underwent serial venipuncture every 3-5days starting in the early follicular phase and lasting up to 10 collections. AMH was tested using the Access 2 immunoassay system. Age-adjusted mixed-effect models utilizing data from 384 samples from 50 subjects demonstrated a within subject standard deviation of 0.81 (95% CI 0.75-0.88) with a coefficient of variation of 23.8% across the menstrual cycle and between subject standard deviation of 2.56 (95% CI 2.13-3.21) with a coefficient of variation of 75.1%. Intra-class correlation (ICC) of AMH across the menstrual cycle was 0.91. Overall, AMH levels, using the automated Access AMH assay, appear to be relatively stable across the menstrual cycle. Fluctuations, if any, appear to be small, and therefore, clinicians may advise patients to have AMH levels drawn at any time in the cycle.

The numbers of 2- 5 and 6-9 mm ovarian follicles are inversely correlated in both normal women and in polycystic ovary syndrome patients: what is the missing link?

Is the negative correlation between the numbers of 2-5 and 6-9 mm follicles influenced by ovarian and/or metabolic parameter(s) in young control women and in patients with polycystic ovarian syndrome (PCOS)? Our study confirmed that the negative correlation between numbers of follicles sized 2-5 and 6-9 mm was stronger in PCOS than in young control women and was not linked to any ovarian or metabolic parameter. Previous reports described a direct negative correlation between the number of small antral follicles (2-5 mm) and large antral follicle (6-9 mm) during the early follicular phase (cycle Days 2-5) in normal and PCOS women. Numerous factors, that could be either intrinsic to the ovary or secondary to metabolic influence and/or gonadotropin regulation, might account for this. Six hundred and thirty-nine patients with PCOS according to Rotterdam Criteria and 157 control women were recruited in this retrospective cross-sectional study from January 2009 to January 2016. Data were obtained from a database of clinical, hormonal and ultrasound (U/S) features recorded consecutively in a single reproductive medicine centre. Univariate correlations between the various parameters were analysed by the Spearman's correlation test. All variables significantly related to the 2-5 and/or 6-9 mm follicle numbers were included in a principal component analysis (PCA) in order to structure the data and to obtain collections of uncorrelated variables, called principal components (PC), which are linear combinations of the original variables. By univariate analysis, the 2-5 and 6-9 mm follicle numbers were strongly but negatively correlated in both populations. Many other variables were correlated to the 2-5 and/or 6-9 mm follicle numbers and to each other. By PCA, these relationships were gathered into four independent PCs in each population. In both groups, the 2-5 and 6-9 mm follicle numbers correlated strongly and inversely to a specific PC. Among the other variables tested, only serum oestradiol level correlated weakly to this PC in the control group. Two other uncorrelated PCs gathered relationships between variables linked to the metabolic status and the gonadotropin regulation both in control and PCOS women. Lastly, a fourth PC included relationships which linked to ovarian ageing in controls and to follicle dysregulation in patients with PCOS. Our controls did not represent the general population since they were recruited in an ART centre; we used a modified Rotterdam classification for PCOS using follicle count and/or serum AMH level with in-house thresholds to define the follicle excess; the AMH assay used is no longer commercially available. Factor(s) regulating specifically the equilibrium between the 2-5 and 6-9 mm follicle numbers still need(s) to be identified. More attention should be paid to the oocyte. None.

Multi-center evaluation of the access amh assay to measure amh as an aid in the prediction of poor ovarian response to controlled ovarian stimulation

Multi-center evaluation of the access amh assay to measure amh as an aid in the prediction of poor ovarian response to controlled ovarian stimulation

What are the likely IVF/ICSI outcomes if there is a discrepancy between serum AMH and FSH levels? A multicenter retrospective study

ObjectiveThe principal outcome was to assess the ovarian response to controlled hyperstimulation during in vitro fertilization (IVF) with or without micro-injection (ICSI) in patients whom ovarian reserve testing revealed a discrepancy between the serum levels of FSH and AMH. The secondary outcome was to determine whether AMH and FSH profiles could predict the IVF/ICSI response. Study designThis was a multicenter, retrospective study analysing all controlled ovarian hyperstimulation cycles with attempted fresh embryo transfer(s) carried out during IVF/ICSI treatment and in which the AMH level had been assayed between January 01, 2008 and December 31, 2011. This enabled us to form 2 control groups (NOR, normal ovarian reserve: normal AMH and FSH and DOR, diminished ovarian reserve: diminished AMH, increased FSH) and 2 study groups (DAMH: diminished AMH, normal FSH and NAMH: normal AMH, increased FSH). The principal assessment criterion was quantitative ovarian response to stimulation defined by the mean number of oocytes punctured, the secondary assessment criterion the qualitative response to stimulation defined by the pregnancy rate per cycle. ResultsWe were able to analyse 1803 stimulation cycles. The mean number of oocytes punctured was significantly reduced in the DAMH and DOR groups compared to the NAMH and NOR groups (5.2±3.9 and 4.1±3.3 vs. 11.5±7 and 9.5±5.6, respectively [P<0.01]). The pregnancy rate per initiated cycle was significantly reduced in the DAMH and DOR groups compared to the NAMH and NOR groups (20% and 24% vs. 32 and 35%, respectively [P<0.01]). Live birth rates did not differ between the groups however. Multivariate analysis with logistic regression revealed that AMH, FSH and age independently had an effect on the number of oocytes punctured, although the effect exerted by AMH seemed to be preponderant (OR: 2.75: 95%CI [2.39–3.19]). AMH appeared to be the sole factor independently predictive of pregnancy per cycle. ConclusionThe serum AMH level appears to provide an additional item of discriminatory information, which should not be overlooked. Ovarian reserve work-up should include routine AMH assay.