High-efficiency chemiluminescent POCT assay for anti-Müllerian hormone: innovative development and application in polycystic ovary syndrome diagnosis

Abstract

In recent years, the decline in ovarian reserve function has become a major cause of reduced female fertility. Early identification, evaluation, and management of ovarian reserve dysfunction are essential for improving female fertility and reproductive health. Currently, the main diagnostic method is assessing ovarian function through measuring AMH serum concentration and follicle count. Traditional ELISA methods for AMH detection are limited by low sensitivity, long detection times, and susceptibility to manual operation errors. Additionally, many existing AMH measurement techniques require expensive equipment and involve complex procedures, making them less suitable for rapid and accessible clinical use. This study developed a chemiluminescence immunoassay (CLIA) method based on a double-antibody sandwich format for POCT of human serum AMH. A fully automated chemiluminescence immunoassay analyzer is developed independently by our laboratory, without the need for an excitation light source, so that it can be measured on the spot. After incubating the sample with the detection antibody, horseradish peroxidase emits light under substrate catalysis. AMH concentration is detected with a fully automated CLIA instrument, offering high sensitivity, micro-volume, efficiency, and low-cost quantitative POCT detection. Under optimal conditions, the detection limit is 0.02 ng mL⁻¹, with a linear range of 0.02–25 ng mL⁻¹ and a detection time of 35 minutes. The method demonstrates excellent stability, reproducibility, and selectivity. Evaluation of 134 clinical serum samples showed a correlation coefficient (R²) of 0.9844 with a Beckman CLIA kit. For diagnosing PCOS, the combined detection of DHEAS, LH, and TESTO resulted in an AUC of 0.99. This POCT method offers a fast, reliable tool for assessing AMH levels and improving ovarian reserve evaluation.