Background: Cytomegalovirus (CMV) infection is the leading non-genetic cause of sensorineural hearing loss. However, there is no standardized protocol for screening newborns for this infection. Objectives: To characterize the CMV testing pattern of neonates born at our institution, and to determine the prevalence of CMV testing in neonates who failed the routine hearing screen. Study Design: We conducted a retrospective chart review for the time period 1/1/2010 through 12/31/2014. Demographics, clinical presentation, and CMV and hearing test results were analyzed. A survey of CMV test indications among neonatology practitioners was conducted. Results: Of 31,016 newborns, we identified a lower than expected rate of symptomatic congenital CMV infection (9 [0.03%] observed vs. 21 [0.7%] expected from general newborn prevalence). 232 infants were screened for CMV; the most common clinical presentations for CMV positive infants were low birth weight, thrombocytopenia, rash and unconjugated hyperbilirubinemia. 37.5% neonates who were CMV positive failed the initial hearing screen compared to 6.2% who were CMV negative. 6.7% of all neonates failed routine newborn hearing screen; only 0.7% of these were tested for CMV. The neonatology practitioner survey showed that the most consistent indications for CMV testing were microcephaly and low birth weight but there was no consistent pattern. Conclusion: Identification of CMV infection was lower than expected; it is likely that mild or only single symptom may be missed as identifiers of CMV. The practices for CMV screening varied widely among clinicians. Newborns who failed routine hearing test were rarely tested for CMV. Development of standardized guidelines for CMV testing is needed.