In recent years, more attention has been put on non-invasive sampling method for monitoring physiological status of wildlife. In this article, we reviewed factors of animal fecal steroid hormone detection accuracy during the detection, hoping to provide a reference for the accurate application of fecal steroid hormone detection technology in wildlife. The factors include the freshness, volume and storage of the sample, daily rhythm and seasonal variation of hormone metabolism, age, sex and reproductive state of animals, etc.