Evaluation of a novel, minimally invasive test for fertility hormones

Abstract

To develop and validate an at-home, self collection method for measuring eight reproductive hormones using minimally invasive fingerstick (FS) sampling on filter paper. A prospective concordance study compared matched venipuncture and FS samples from 130 women ages 18 to 40 on menstrual cycle day three. All women provided written, informed consent. Precision, linearity, and accuracy studies were also conducted. Samples were measured for anti-Müllerian hormone (AMH), estradiol, (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), testosterone (T), thyroid stimulating hormone (TSH), and free thyroxine (FT4). One-hundred thirty matched venipuncture and FS samples were tested using FDA-cleared immunoassays, with a modified reconstitution step for FS samples. Concordance was evaluated in the clinical samples, in addition to six matched contrived samples to ensure linearity and coverage over the assay range. Deming regression lines were fit, and bias was assessed using Bland-Altman plots. We compared our bias calculations with the optimal limits established by the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM). Linear relationships were assessed by Pearson product-moment correlation coefficients. Precision was calculated by measuring 60 repeats of high and low controls for each assay. Accuracy/percent recovery was calculated by comparing triplicate measures of five levels of manufacturer-supplied calibrators. Venipuncture and FS hormone values were highly concordant and linear across all assay ranges (Table 1). Bias measures were uniform and below EFLM biases. The correlation between venipuncture and FS results was 0.99 for each hormone. Each assay displayed a high degree of precision with less than 15% coefficient of variation. Each assay displayed a high level of accuracy (average recovery ranging from 95.5% to 102.3%). Recovery was within the 15% limit of acceptability for biological analyses, as recommended by the FDA. Venipuncture and FS samples can be used interchangeably to measure the following hormones in women: AMH, E2, FSH, LH, PRL, T, TSH, FT4. Fingerstick sampling will allow women to test reproductive hormone levels at home--giving both women trying to conceive and women planning for future fertility the ability to take ownership over their reproductive options. These hormone tests may also help screen for conditions such as polycystic ovarian syndrome or primary ovarian insufficiency.Tabled 1Concordance results between venipuncture and fingerstick samplesHormoneDeming Regression EquationCorrelation Coefficient (*p<.0001)Average Percent BiasAMH (ng/mL)Y = 1.02*X - 0.040.99*6.72%E2 (pg/mL)Y = 1.03*X - 2.030.99*-5.07%FSH (mIU/mL)Y = 1.00*X + 0.120.99*1.14%LH (mIU/mL)Y = 1.09*X - 0.270.99*2.75%PRL (ng/mL)Y = 1.03*X - 0.060.99*2.93%T (ng/mL)Y = 1.00*X0.99*-1.87%TSH (uIU/mL)Y = 1.02*X - 0.030.99*0.58%FT4 (ng/dL)Y = 0.99*X + 0.020.99*1.38% Open table in a new tab