Background: Otoacoustic emissions (OAEs) and auto-auditory brainstem response (AABR), as two safe and equally accurate techniques, are used for hearing screening among newborns. However, the screening time of such tests is under debate. Objectives: The present study aimed to assess the correlation between examination day and the referral rate of secondary hearing screening among non-high-risk newborns. Methods: A retrospective review of secondary hearing screen data collected from June 2012 to June 2019 was conducted on infants who had no confirmed risk factors introduced by the Joint Committee of Infant Hearing 2007 (JCIH). Results: Of the 2493 newborns included in this study, 2129 cases (85.4%) passed the test bilaterally, and 364 newborns (14.6%) failed the examination. The referral rate of the 1366 newborns taking OAE was 13.1%. Among 1127 newborns taking both OAE and AABR, the referral rate was 16.5%. Moreover, the referral rate of the OAE and OAE+AABR techniques was the lowest in the 42-56-day group. Conclusions: All newborns with no high-risk factors should be screened for hearing as such we recommend 42 - 56 days after birth as the best re-examination period to reduce the false positive rate and caregivers' anxiety.