Validation of a new multiplex assay against individual immunoassays for the quantification of reproductive, stress, and energetic metabolism biomarkers in urine specimens

Abstract

Measuring multiple hormones simultaneously in a single assay saves sample volume, labor, time, reagents, money, and consumables. Thus, multiplex arrays represent a faster, more economically and ecologically sound alternative to singleton assays. To validate a new, commercially available multiplex female array produced by Quansys Biosciences against individual immunoassays for the quantification of six hormones in urine samples from women in different reproductive stages. Urine samples were analyzed using the new Quansys multiplex female hormone array and compared with well-established individual immunoassays for adiponectin, free cortisol, c-peptide, estrone-3-glucuronide (E₁G), follicle stimulating hormone beta-subunit (FSH-beta), and human chorionic gonadotropin beta-subunit (hCG-beta). Correlations between assays were assessed using Pearson correlation, linear regression and Bland-Altman analysis. The temporal profiles of free cortisol, E1G, FSH-beta, and hCG-beta were also compared. The multiplex array was highly correlated with the individual immunoassays for five of the tested hormones (Pearson's correlation coefficient ≥ 0.75), and yielded temporal patterns of hormone profiles consistent with the individual immunoassays for free cortisol, E₁G, FSH-beta, and hCG-beta. The Quansys multiplex female hormone array is a valid alternative method to individual immunoassays for the quantification of stress, reproductive and energetic hormones and metabolites in human urine samples and can be used to examine the dynamic interactions between these hormones.