Abstract Study question Do SpermQT results prospectively correlate with fertility outcomes of couples undergoing intrauterine insemination (IUI)? Summary answer We observe a decrease in IUI pregnancy rates in men with Abnormal compared to Normal SpermQT results irrespective of their sperm total motile count (TMC). What is known already As the mainstay of male infertility diagnosis, the standard semen analysis provides limited value for predicting pregnancy outcomes. New technologies are needed for a more comprehensive assessment of male fertility. A novel, epigenetic assessment of sperm quality, SpermQT, was implemented clinically in 2022. Retrospective studies have shown that sperm health, as identified through DNA methylation patterns, is predictive of IUI outcomes in a subset of infertile men. Additional prospective research is needed for validation of these data. Study design, size, duration Over an 18-month period (June 2022 - January 2024), 23 reproductive endocrinologists over eight clinics ordered 488 SpermQT tests on male patients. For these patients, physicians and clinical support staff were asked to provide the sperm total motile count (TMC) from the semen analysis, male and female ages, procedure information and outcomes. SpermQT results were categorized as Normal or Abnormal. Associations between SpermQT results, outcomes, age, and TMC were analyzed. Participants/materials, setting, methods The selection of patients for SpermQT assessment was based on the clinical decision of the ordering physician. SpermQT requires a semen sample which was collected by the patient either at the andrology laboratory or at home using an at-home collection kit. All sample types were shipped directly to the vendor’s laboratory for analysis and report generation. Following the completion of the epigenetic analysis, all SpermQT results were provided to the ordering physician for patient consultation. Main results and the role of chance 488 patients received a SpermQT result. 83.6% of men had a Normal SpermQT (n = 408), and 16.4% had an Abnormal SpermQT (n = 80). 180 couples had pregnancy outcomes from at least one IUI. Of the 180, 14% of men had an Abnormal SpermQT (n = 25) and 86% had a Normal SpermQT (n = 155). Analysis of pregnancy outcomes from all 180 couples who completed one, two, or three IUI cycles showed a 15% IUI cumulative pregnancy rate. The men with a Normal SpermQT had a 17.42% cumulative IUI pregnancy rate within the first three IUI cycles while men with an Abnormal SpermQT (N = 25) had no reported pregnancies and a 0% cumulative IUI pregnancy rate. Within this patient cohort, 338 patients had reported TMC where 82% had a TMC ≥20 million (n = 276). 72% of men with an Abnormal SpermQT had a TMC ≥20M, while 84% of men with a Normal SpermQT had a TMC ≥20M. Median male and female ages in the Normal SpermQT category were both 34, while median male and female ages in the Abnormal SpermQT category were 32 and 33 respectively, indicating age did not play a role in IUI pregnancy outcomes. Limitations, reasons for caution Limitations within this study include the sample size of men with an Abnormal SpermQT result. In order to identify a statistical significance between IUI pregnancy outcomes, a total sample size of 500 patients will be needed. Data and outcomes will continue to be collected to continue the validation of SpermQT. Wider implications of the findings There is strong data showing that sperm epigenetics play an important role in IUI outcomes. Here we show in a prospective and multi-site population that a new assessment of sperm epigenetics (SpermQT) identifies men less likely to achieve an IUI pregnancy, and also identifies subfertile men with normal TMC. Trial registration number Non Applicable