The Royal College of Pathologists of Australasia's Quality Assurance Programs (QAP) on serologic assays for infectious Mononucleosis (IM) have identified a number of important problems in laboratory diagnosis of this condition. A wide range of assays for the diagnosis of acute Epstein-Barr virus (EBV) infection are available, although heterophile antibody tests are still the most frequently performed procedures for the diagnosis of IM. In 1993, eighty-six (54%) of the 159 participating laboratories performed only a heterophile test; of these, 71% did not provide an interpretation of their results and none mentioned the need for confirmatory EBV-specific antibody testing. This revealed a lack of appreciation that heterophile tests should only be used for screening, due to their inferior sensitivity of less than 50% in children and 80-90% in adults and specificity of 95% compared to EBV-specific assays. For these reasons the use of heterophile tests is discouraged. Although EBV-specific serology has traditionally been done by immunofluorescence (IF), the use of reliable ELISA methods using purified EBV-antigens is increasing. False negative EBV VCA IgM ELISA test results were obtained by laboratories using unpurified or unspecified VCA antigens. As only five laboratories used these tests in the 1992 QAP, the tests' true performances could not be properly assessed, suggesting the need for independent studies. Variable results were obtained by laboratories using commercial EBNA IgG assays (in addition to EBV IgM) suggesting that an assessment of these kits would also be worthwhile. The QAP reaffirm that reliable EBV IgM detection is the most useful test for the diagnosis of IM, even where other EBV markers are also used. In the 1992 and 1993 QAP, laboratory performance of both heterophile and EBV-specific tests was better with non-reactive serum specimens (99% (correct) than with reactive serum specimens (90% correct).