Inter-assay variation and reproducibility of progesterone measurements during ovarian stimulation for IVF

Abstract

In recent years there is increasing evidence that elevated progesterone levels during ovarian stimulation for IVF / ICSI have a negative impact on the ART-outcome. However, different progesterone assays were used in the previous studies and different assays might produce varying results. This retrospective study evaluated the reproducibility and reliability of different progesterone assays with a special focus on progesterone levels below 1.5 ng/ml, as this range is crucial for early detection of progesterone rise during ovarian stimulation for IVF. A total of 413 blood samples were categorized in different progesterone ranges and whether they were retrieved on the day of final oocyte maturation and the results were compared regarding their reproducibility and reliability. To compare the reproducibility between the different progesterone assays, the Intraclass Correlation Coefficient (ICC) was calculated and interpretation of the ICC results was done according to Cicchetti, ranging from poor to excellent. The correlation of the assays was excellent when all samples were compared including samples retrieved on day of final oocyte maturation, however in the ranges of progesterone levels 1.0 ng/ml to < 1.5 ng/ml, 0.8 ng/ml to < 1.0 ng/ml and < 0.8 ng/ml, the ICC varied between poor and excellent. The assays “gen III” and “Architect” showed an excellent reproducibility of progesterone results throughout all ranges of progesterone levels.This analysis demonstrates, that different progesterone assays have a limited reproducibility and that the results depend on the assay used and the range of progesterone level. This fact leads to two important conclusions. Firstly the limited reproducibility might lead to substantially different treatment decisions in ovarian stimulation treatment for IVF and secondly critical interpretation of thresholds, provided by meta-analysis, is crucial despite the risk that the so far gained clinical experience might become irrelevant and has to be adjusted to the results, obtained by each assay.

Longitudinal analysis of external quality assessment of immunoassay-based steroid hormone measurement indicates potential for improvement in standardization

“…Overall, the bias analysis of the testosterone, progesterone and 17β-estradiol data confirmed the findings of previously published studies, which found that immunoassays were insufficiently reliable in quantitatively determining sex hormones. A trend towards standardizing immunoassay detection for steroids has yet to be observed ( Vesper et al, 2014 ; Lawrenz et al, 2018 ). However, this EQA data revealed one exception.…”

Low Luteal Serum Progesterone Levels Are Associated With Lower Ongoing Pregnancy and Live Birth Rates in ART: Systematic Review and Meta-Analyses

“…However, it is important to keep in mind the main pitfalls in determining optimal serum progesterone levels. Serum progesterone levels vary according to the assay applied for its determination (48,49). Hence, each clinic might need to determine its own threshold.…”

Impact of Elevated Progesterone in Late Follicular Phase on Early Pregnancy Outcomes and Live Birth Rate After Fresh Embryo Transfers

“…Our study is the first known study to quantify the relationship between progesterone elevation in the late follicular phase and pregnancy outcomes in the IVF fresh cycle by smoothing curve fitting and threshold effect analysis. An observational retrospective study demonstrated that different progesterone assays had a limited reproducibility and results depending on the specific assay used and the range of progesterone level ( Lawrenz et al, 2018 ). Consequently, the reliability of progesterone thresholds, suggested by a previously published meta-analysis on the topic ( Venetis et al, 2013 ), has to be questioned because studies using different assays were included in that meta-analysis.…”