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Improving retention in care is a key element of the National HIV/AIDS Strategy (NHAS). However, definitions for measuring retention in care are not standardized.To compare measures of retention based on both clinic visit data and HIV laboratory surveillance data.Retrospective cohort study.New York City (NYC), New York.We matched adult patients with HIV infection seen at the Spencer Cox Center for Health (SCC) in 2010 or 2011 with the NYC HIV Surveillance Registry.Retention in care was measured on the basis of SCC electronic medical record (EMR) data (≥1 medical visits in 2012) and Surveillance Registry data (≥2 CD4/viral load [VL] tests ≥90 days apart in 2012).There were 5746 adult HIV-infected patients seen at SCC between 2010 and 2011 who matched with the Surveillance Registry. Seventy-eight percent (n = 4469) had 1 or more medical visits at SCC in 2012 and were considered retained on the basis of the EMR definition, among which 3831 (86%) met the surveillance definition for retention in care. Patients who did not have a medical visit at SCC in 2012 (n = 1277) were lost to care in NYC (n = 485; 36%), engaged in care at an alternate provider (n = 622; 49%), or died after their last SCC visit (n = 197; 15%).This study is an important comparison of laboratory surveillance versus clinic visit-based measures of retention in care in an urban setting with the largest HIV epidemic in the country. Collaborative projects between local health departments and clinical care providers can help validate the care status of patients and inform the allocation of resources to reengage patients who are lost to care.The combined use of laboratory and clinic visit–based data to measure retention in care provides a more accurate representation of the care status of HIV-infected patients than use of a single data source alone. Routine sharing of data by public health institutions and clinical care providers would help target resources toward reengaging patients who are lost to care in jurisdictions with universal HIV-related laboratory reporting.