In the application of assisted reproductive technology (ART), the impact of antinuclear antibody (ANA) on the pregnancy outcomes of infertile patients has garnered attention. ANA, as a biomarker of autoimmune abnormalities, has been extensively studied for its association with unexplained infertility and its potential effects on oocyte and embryo quality. Current evidence suggests that a positive ANA status may be associated with reduced ovarian reserve function, poor embryo development, decreased clinical pregnancy rates, and increased risk of miscarriage. The level of ANA in follicular fluid is considered a sensitive indicator for predicting autoimmunity-related infertility. Specific ANA subtypes, such as anti-dsDNA antibodies and anti-centromere antibodies (ACA), are related to oocyte maturation disorders and delayed embryo development. Nevertheless, the direct link between a positive ANA status and the decline in female fertility and adverse reproductive outcomes requires further research for clarification. The review also discusses various treatment options for ANA-positive patients, including glucocorticoids, low-dose aspirin, and hydroxychloroquine, aiming to provide guidance for clinical practice in ART and future research directions.