The standard for diagnosing meningoencephalitis includes cerebrospinal fluid (CSF) culture and viral polymerase chain reaction (PCR). Approval of the FilmArray® BioFire® Meningitis/Encephalitis (ME) panel has reduced time to detection of several pathogens and improved diagnostic sensitivity. The objective of this study was to determine the impact on intravenous (IV) acyclovir duration of the ME panel compared to previously utilized CSF studies within a large health system with a central laboratory. A multicenter quasi-experimental cohort study of adult and pediatric patients was conducted (n = 208). The primary endpoint was duration of IV acyclovir, which was decreased (41.6 v. 30.8 hours; P < 0.01) with the ME panel. Secondary outcomes including test-turnaround time (TAT) and the impact of utilizing a central laboratory were explored. Subgroup analyses demonstrated that number of daily couriers from hospital to the central laboratory (0 versus 7 versus 3 versus 2 couriers) and hospital distance from the central laboratory (0 versus 1–10 versus 11–20 versus 21–30 miles) significantly impacted TAT (P < 0.01). While duration of IV acyclovir for the entire healthcare system was reduced, the duration at individual sites was not impacted by number of couriers or distance from the central laboratory.