Originally, NVSL was part of the Bureau of Animal Industry. 

In 1961, National Animal Disease Laboratory (NADL) opened in Ames, Iowa with a Director, an Assistant Director for Research, and an Assistant Director for Regulatory Laboratories.  Regulatory Laboratories occupied twenty percent of NADL space and provided diagnostic services for the Animal Disease Eradication Division and biologics evaluations for the Animal Inspection and Quarantine Division. 

A few years later, reorganization resulted in three rather independent units: research, biologics and diagnostics. The biologics group was located outside of NADL facilities.  In 1971, Animal Health Division (AHD) laboratory facilities in Beltsville, Maryland, were assigned to the Diagnostic Services group.

In 1973, Biologics and Diagnostic Services Laboratories were brought back together under one Director and named Veterinary Services Laboratories.

In December 1977, the name changed to National Veterinary Services Laboratories (NVSL).  Plans were made for construction of new facilities as growth continued.  Phase 1 of the plan was completed in 1978 and the biologics, administrative and support functions moved into the new building.  That year, Animal and Plant Health Inspection Service, (formerly AHD), located in Beltsville, Maryland, closed and the functions moved to Ames.

In 1984, diagnostic activities at Plum Island Animal Disease Center located in Plum Island, New York, were transferred to APHIS' supervision.  It was given the name Foreign Animal Disease Diagnostic Laboratory and was part of NVSL.

Biologics testing activities split from NVSL and joined the inspection and licensing activities in forming the Center for Veterinary Biologics in 1996.  The NVSL now focuses exclusively on diagnostic services.

APHIS and ARS are building a modernized and consolidated facility for animal health research, diagnosis, and product evaluation for NADC, NVSL, and CVB.  Building 21 (Phase 1) was completed September 13, 2004 and houses Pathobiology and Diagnostic Bacteriology Laboratories.  Phase 2-the High Contaminant Large Animal Facility (BSL-3Ag), the Low Containment Large Animal Facility (BSL-2Ag), and the Consolidated Laboratory Facility (which will contain all the remaining laboratory/office/support space), are all projected to be completed by the end of 2009.