Comparison of Western immunoblotting and microimmunofluorescence for diagnosis of Mediterranean spotted fever

Abstract

One-hundred serum samples from 41 patients suffering from Mediterranean spotted fever (MSF) were tested by microimmunofluorescence (MIF) and Western blot (WB; immunoblot). Immunoglobulin G (IgG), IgM, and IgA antibody-specific responses to the high-molecular-mass species-specific protein antigens (115 kDa and 135 kDa) of Rickettsia conorii, as well as to cross-reactive lipopolysaccharide (LPS) antigens, were observed. The WB assay detected IgM-type antibodies earlier than did the MIF assay. These antibodies were often directed against nonspecific LPS and may have a questionable positive predictive value. In addition, an IgG reaction to a 60-kDa protein was observed in four cases of malignant forms of MSF but was never observed in cases of mild forms. This reaction could be correlated with a marker of the severity of the development of MSF. From a previous MIF survey of blood donors, 9 negative, 11 IgG-positive, and 6 IgM-positive serum samples were selected for comparison by WB. Sera negative by MIF were also negative by WB. MIF IgG-positive sera showed a specific response to R. conorii in the WB assay, but the six serum samples from this seroepidemiological study positive for IgM by MIF were almost all negative by the WB assay. One was positive for IgM against the LPS but was considered a false positive. The WB is shown to provide a new tool for serodiagnosis.