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Tissue safety and added testing is a frequent topic of discussion among those who must evaluate donor suitability and assure appropriate testing. Therefore, the purpose of this report was to examine the data of one eye bank (accredited by Eye Bank Association of America) for the presence or absence of reactive serology in 3592 donors over a 5-year period, from 2005 to 2010.The number of specific analytes, human immunodeficiency virus (HIV), hepatitis B virus, or hepatitis C virus, that were reactive and the number of multireactive serologies are presented along with reports related to the identification of nucleic acid testing (NAT), NAT-reactive donors in correlation with other antigen or antibody markers. Antigen or antibody markers are enzyme-linked immunosorbent assay immunoassay kit determinations. NATs are used to measure the presence of RNA or DNA virus.The 249 donors with reactive serologies serve to confirm the importance of serologic testing in history-negative potential donors. Furthermore, the findings of 2 HIV NAT-only reactive individuals is significant because it may indicate an early unrecognized exposure and “window period” infectivity not recognized by antibody testing alone. These results support the appropriate inclusion of this added test to guarantee tissue safety.The finding of 2 potential donors without identified risk factors but with reactive HIV NAT suggests the need for comprehensive serologic testing in conjunction with medical social history screening. Considered together, screening and testing offer the best allograft tissue safety currently available.