Soluble LH-HCG receptor and oestradiol as predictors of pregnancy and live birth in IVF.

RESEARCH QUESTION: Circulating soluble LH-HCG receptor (sLHCGR) is a first-trimester marker for screening pregnancy pathologies and predicts premature or multiple births before fertility treatment. Oestradiol per oocyte at ovulation induction predicts IVF treatment outcomes. We asked whether sLHCGR levels are stable during fertility treatment and whether, alone or with oestradiol, they could improve prediction of fertility treatment outcomes. DESIGN: Serum sLHCGR, anti-Müllerian hormone [AMH] and oestradiol were measured in patients undergoing IVF. Antral follicle count before ovarian stimulation and oocyte yield were used to establish sLHCGR- oocyte ratio (SOR), sLHCGR- antral follicle ratio (SAR), oestradiol at trigger per oocyte (oestradiol-oocyte ratio [EOR]) and oestradiol at trigger per antral follicle (oestradiol-antral follicle ratio [EAR]). RESULTS: The relatively stable sLHCGR was negatively related to AMH when oocyte yield was high. The sLHCGR levels were proportional (r = 0.49) to oestradiol at early cycle (day-3). Pregnancy and live birth were highest at low sLHCGR (≤1.0 pmol/ml) and SOR (≤ 0.1 pmol/ml/oocyte). A total of 86-89% of live births in IVF treatment were within the cut-off parameters of SAR and SOR (0.5 pmol/ml) and EAR and EOR (380 pg/ml). For failed pregnancy, age, SOR and EOR together had positive and negative predictive values of 0.841 and 0.703, respectively. CONCLUSIONS: sLHCGR levels are negatively related to AMH when oocyte yield is high. High early cycle sLHCGR is associated with elevated day-3 oestradiol. Low sLHCGR and SOR are indicators of increased clinical pregnancy and live birth rates. Patient age and SOR, combined with EOR, might improve prediction of IVF treatment outcomes.

Objective multicentre performance of the automated assays for AMH and estimation of established critical concentrations.

The measurement of AMH has now become widespread practice within the field of fertility treatment and research, despite technical issues with some of the original assays. The two new automated assays, with their potentially improved technical performance, require detailed examination and comparison under different conditions. In addition, the determination of categories of responses to ovarian stimulation, require re-evaluation for these new tests. The performance of the assays across numerous laboratories, and over a protracted timeframe, has been examined through the UK NEQAS published results. The automated assays show high quality performance figures over a broad concentration range, with exceptionally low variance figures, and they also yield very similar absolute concentration values. Critical response diagnostic concentrations have been re-evaluated by determination of age-related concentrations from within large population datasets.

Premature and multiple births in IVF are associated with pretreatment circulating LH/hCG receptor concentration.

The luteinizing hormone (LH) and pregnancy hormone, human chorionic gonadotrophin (hCG), share a common receptor: LH/hCG-R or LHCGR. In this prospective study involving 290 patients undergoing in vitro fertilization (IVF) and embryo transfer, we have examined whether pretreatment circulating LHCGR (sLHCGR) influences the course of pregnancy and perinatal outcome after embryo transfer. The blood samples were collected before the fertility treatment began and sLHCGR concentrations were measured using an enzyme-linked immunosorbent assay (ELISA) test. We demonstrate that extreme pretreatment sLHCGR concentrations (low & high) were linked to abnormal birth weights for singleton births, while very low concentrations of sLHCGR were associated with premature delivery (≤34 weeks) of singletons and multiple births following transfer of ≥2 embryos.

Stability of AMH measurement in blood and avoidance of proteolytic changes.

The new Gen II assay for anti-Müllerian hormone (AMH) shows good stability and reliability in serum, but analyses of stability in whole blood are lacking. Testing the effects of storage of whole-blood samples at room temperature revealed significant increases in the measured value of AMH of 31% over 4 days (P<0.001). The effect is temperature dependent, with storage at 4°C showing markedly reduced increments. Further, samples collected into serum tubes with gel separators and centrifuged within 5h (blood cells and serum physically separated within the collection tube) showed reliable stability over a period of more than 5 days.