In vitro and in vivo assays have been used for the detection of reticuloendotheliosis virus (REV) in live virus vaccines of poultry. The presence of REV is confirmed by the demonstration of viral antigen or provirus in chicken embryo fibroblasts (CEFs) or in specific-pathogen-free chickens inoculated with vaccine. Using REV polyclonal or monoclonal antibodies, CEFs inoculated with vaccines can be examined for REV by immunofluorescence or immunoperoxidase staining methods. Cell lysates from such inoculated CEFs can also be used for detection of REV major group-specific antigen (p30) by an enzyme-linked immunoassay. Detection of proviral DNA by polymerase chain reaction (PCR) assays that amplifies the 291 base pairs product of REV LTR has been shown to be a sensitive and specific method for detection of various strains of REV in infected CEFs and in the blood of SPF chickens inoculated with contaminated fowlpox virus (FPV) vaccines. Recently, using PCR tests that amplify REV envelope and REV 3' LTR sequences provided a more accurate assessment of the insertion of REV provirus in FPV than PCR assays that amplify the REV 5' LTR. This paper reviews the most common methods used for testing live virus vaccines of poultry for contamination with REV.