Aspects of the diagnosis of Lyme borreliosis.
Attempts were made to culture spirochetes from the cerebrospinal fluid of 105 patients with suspected Lyme borreliosis with neurological complications. At the final evaluation, only 38 patients fulfilled the criteria of neuroborreliosis. Spirochetes were cultured from the cerebrospinal fluid of four of these patients. All four had pleocytosis in their cerebrospinal fluid and a history of neurological symptoms of only four to ten days. Two had no detectable antibodies in their cerebrospinal fluid against any of the isolated spirochetes, neither when tested with an ELISA nor by Western blot. A distinctly stronger antibody reaction to the homologous isolate than to the heterologous isolates was found in serum and cerebrospinal fluid from one patient. The cells of the isolates were morphologically similar and showed a very similar protein pattern when analyzed by SDS-PAGE. Cells of all isolates reacted with the monoclonal antibodies H5332 and H9724, which also react with Borrelia burgdorferi isolate B31, the type species. One isolate lost a major protein of 23 kD after subcultivation for four months. We conclude that isolation of spirochetes from cerebrospinal fluid is not suitable as a routine method but might prove successful in clinically selected cases of Lyme borreliosis. The patient antibody response to spirochetal components was analyzed with Western blot. Antibodies to low-molecular components including a major protein with a molecular weight of 21-23 kD, and to a 41-kD major flagellar protein, were the first to appear in serum and in CSF samples. No single band in the immunoblots was found to be specific. By requiring a 41 kD band together with at least one low-molecular band for a positive immunoblot, 53 of 68 (78%) patients with neuroborreliosis had positive IgM and/or IgG serum immunoblots by visual reading of coded material. Western blot was more sensitive than ELISA based on a sonicate antigen which identified 40 of the 68 (59%) patient samples as positive, but not significantly more sensitive than ELISA based on a purified flagellum antigen which identified 50 of 68 (74%). Western blot tended to be more sensitive than the flagellum ELISA regarding sera from patients with neurological symptoms of 2 weeks or shorter duration. However, there was a tendency towards a lower specificity regarding the serological diagnosis of current Lyme borreliosis by Western blot than by sonicate and flagellum ELISAs.(ABSTRACT TRUNCATED AT 400 WORDS)
Karlsson
M
M
Department of Infectious Diseases, Danderyd Hospital, Stockholm, Sweden.
D000328:Adult; D000907:Antibodies, Bacterial / Q000096:biosynthesis / Q000134:cerebrospinal fluid; D000911:Antibodies, Monoclonal / Q000276:immunology; D000942:Antigens, Bacterial / Q000032:analysis / Q000276:immunology; D001426:Bacterial Proteins / Q000032:analysis; D015153:Blotting, Western; D015748:Borrelia burgdorferi Group / Q000276:immunology / Q000302:isolation & purification* / Q000648:ultrastructure; D002555:Cerebrospinal Fluid / Q000382:microbiology; D002648:Child; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005407:Flagella / Q000276:immunology; D006801:Humans; D008193:Lyme Disease / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D008297:Male; D008854:Microscopy, Electron; D008875:Middle Aged; D011237:Predictive Value of Tests
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins