Mobile point-of-care diagnostics are paramount for the provision of healthcare. Hormonal diagnostics are powerful tools to monitor timely changes in human physiology. Hormone concentrations in serum directly correlate with urine excretions with minor time delays. Therefore, rapid tests for hormones in urine have been widely used for decades as means of early diagnostics, particularly in lateral flow immunoassay formats. However, the challenge of reading and interpreting these binary tests remains. Here we present a method for utilizing mobile technologies to quantitatively read and interpret hormonal test strips. The method demonstrates the detection of a urinary by-product of progesterone, pregnanediol glucuronide (PdG), and its relation to ovulation and the fertility cycle.

Background: Progesterone is essential for establishing and maintaining a pregnancy. However, premature atresia of the corpus luteum (CL) or sub-optimal progesterone secretion after ovulation may result in the decreased ability to establish or maintain a pregnancy. The current limitations of serum hormone testing fail to give a complete picture of progesterone production across the luteal phase required for pregnancy.Methods: Pregnanediol-3a-glucuronide (PdG) is a progesterone metabolite that can be detected via non-invasive, self-administered urinary test strips and can be used to monitor progesterone production. PdG test strips were used by 185 women and taken multiple times during the luteal phase. Results were analysed by Fisher’s exact test.Results: Of the 185 enrolled women, 172 had complete cycles. Of the 172 women who demonstrated complete cycles, 54 (31.4%) women reported pregnancies. Thirty-five (64.8%) of these pregnancies resulted from a PdG positive cycle with a miscarriage rate of 14.3% (5/35). Nineteen pregnancies (35.2%) resulted from PdG negative cycles with a miscarriage rate of 89.5% (17/19). The association of a negative or positive cycle with pregnancy outcome was significant (two-sided p=0.0001). Positive PdG cycles are associated with increased pregnancy rates and lower odds of a first-trimester pregnancy loss.Conclusions: A consideration for luteal phase support with progesterone or ovulation induction in women with negative PdG cycles can be of benefit to increase pregnancy rates. Furthermore, the use of LH and PdG testing prior to conception can be used as a screening tool to potentially identify women who are at risk of subfertility and at a higher risk of first-trimester pregnancy loss related to low progesterone.Trial registration: NCT05033366. Registered 02 September 2021, https://clinicaltrials.gov/ct2/show/NCT05033366

Background and Objectives: To achieve pregnancy, it is highly beneficial to identify the time of ovulation as well as the greater period of fertile days during which sperm may survive leading up to ovulation. Confirming successful ovulation is also critical to accurately diagnose ovulatory disorders. Ovulation predictor kits, fertility monitors, and tracking apps are all available to assist with detecting ovulation, but often fall short. They may not detect the full fertile window, provide accurate or real-time information, or are simply expensive and impractical. Finally, few over-the-counter products provide information to women about their ovarian reserve and future fertility. Therefore, there is a need for an easy, over-the-counter, at-home quantitative hormone monitoring system that assesses ovarian reserve, predicts the entire fertile window, and can screen for ovulatory disorders. Materials and Methods: Proov Complete is a four-in-one at-home multihormone testing system that utilizes lateral flow assay test strips paired with the free Proov Insight App to guide testing of four hormones—FSH, E1G, LH, and PdG—across the woman’s cycle. In a pilot study, 40 women (including 16 with a fertility-related diagnosis or using fertility treatments) used Complete for one cycle. Results: Here, we demonstrate that Proov Complete can accurately and sensitively predict ovarian reserve, detect up to 6 fertile days and confirm if ovulation was successful, in one easy-to-use kit. Ovulation was confirmed in 38 cycles with a detectable PdG rise. An average of 5.3 fertile days (from E1G rise to PdG rise) were detected, with an average of 2.7 days prior to LH surge. Ovulation was confirmed via PdG rise an average of 2.6 days following the LH surge. While 38/40 women had a PdG rise, only 22 had a sustained PdG level above 5 ug/mL throughout the critical implantation window, indicating ovulatory dysfunction in 16 women. Conclusion: Proov Complete can detect the entire fertile window of up to 6 fertile days and confirm ovulation, while also providing information on ovarian reserve and guidance to clinicians and patients.