We compared the MRL and the Labsystems Chlamydia pneumoniae microimmunofluorescence (MIF) immunoglobulin G (IgG) kits and the Labsystems enzyme immunoassay (EIA) kit in a blinded study of 83 serum samples in which we evaluated titers, cross-reactivity to other species, and reproducibility. There was no statistically significant difference between the MRL and the Labsystems MIF kits in the endpoint titers of IgG antibody to C. pneumoniae. The correlation between the results obtained with these two MIF kits was excellent (r ؍ 0.95; P ؍ 0.001). The cross-reactivity of the C. pneumoniae-positive sera with C. trachomatis-and C. psittaci-positive sera was assessed for each MIF kit. For C. pneumoniae-positive sera with titers of >32, the Labsystems MIF kit exhibited more cross-reactivity to C. psittaci than the MRL kit did While there are no wholly satisfactory serologic methods for the diagnosis of Chlamydia pneumoniae infections, the microimmunofluorescence (MIF) test, when it is properly performed and when its results are properly read, provides the most sensitive and species-specific method for laboratory diagnosis of acute infection (3,8). This test was originally developed by Wang and Grayston (10) in 1970 for detection of C. trachomatis antibodies. The MIF test is an indirect fluorescent-antibody test that measures specific antibodies to epitopes present in the cell walls of the elementary body (EB) particles. Sensitivity and specificity can be improved by using purified EBs of all three species of Chlamydia rather than reticulate bodies, which predominantly express genus-specific epitopes. The MIF test is the only antibody test available that measures the titers of specific antibodies to all species simultaneously. The disadvantages of the MIF test are that the endpoint fluorescence, or the titer, is determined subjectively (8, 11), the test has low throughput, and the test requires proficiency and experience for correct reading of the endpoint titers.Commercially available kits for detection of antibodies to C. pneumoniae are available. MRL and Labsystems both manufacture MIF kits. Cross-reactivity between species is reduced in these kits by treating the EBs to remove genus-specific lipopolysaccharide (LPS). Labsystems also offers an immunoglobulin G (IgG) enzyme immunoassay (EIA) kit (also available in IgA and IgM formats) for detection of antibodies to C. pneumoniae. It could offer a high-throughput alternative to the MIF test if it proves to be specific and sensitive. Reading of its endpoint is objective, and it has the capacity to assay at least 200 serum samples per day. It uses a non-LPS antigen that is reported to be specific for C. pneumoniae. Its disadvantage is that the results are reported as positive or negative enzymeimmuno units (EIU) on the basis of the results for a single dilution of a serum sample and not as an endpoint titer; in addition, when low volumes are to be tested, the EIA plates may in fact become costly due to waste.Commercial kits offer advantages over in-house assays: they sa...