Didier Raoult, M.D., Ph.D., Marie L. Birg, Bernard La Scola, M.D., Ph.D., Pierre E. Fournier, M.D., Ph.D., Maryse Enea, Hubert Lepidi, M.D., Ph.D., Veronique Roux, Ph.D., Jean-Charles Piette, M.D., François Vandenesch, M.D., Ph.D., Denis Vital-Durand, M.D., and Tom J. Marrie, M.D.
Background Whipple's disease is a systemic bacterial infection,but to date no isolate of the bacterium has been establishedin subculture, and no strain of this bacterium has been availablefor study.
Methods Using specimens from the mitral valve of a patient withendocarditis due to Whipple's disease, we isolated and propagateda bacterium by inoculation in a human fibroblast cell line (HEL)with the use of a shell-vial assay. We tested serum samplesfrom our patient, other patients with Whipple's disease, andcontrol subjects for the presence of antibodies to this bacterium.
Results The bacterium of Whipple's disease was grown successfullyin HEL cells, and we established subcultures of the isolate.Indirect immunofluorescence assays showed that the patient'sserum reacted specifically against the bacterium. Seven of 9serum samples from patients with Whipple's disease had IgM antibodytiters of 1:50 or more, as compared with 3 of 40 samples fromthe control subjects (P<0.001). Polyclonal antibodies againstthe bacterium were generated by inoculation of the microorganisminto mice and were used to detect bacteria in the excised cardiactissue from our patient on immunohistochemical analysis. The16S ribosomal RNA gene of the cultured bacterium was identicalto the sequence for Tropheryma whippelii identified previouslyin tissue samples from patients with Whipple's disease. Thestrain we have grown is available in the French National Collection.
Conclusions We cultivated the bacterium of Whipple's disease,detected specific antibodies in tissue from the source patient,and generated specific antibodies in mice to be used in theimmunodetection of the microorganism in tissues. The developmentof a serologic test for Whipple's disease may now be possible.
Source Information
From the Unité des Rickettsies, Université de la Méditerranée, Faculté de Médecine, Marseilles, France (D.R., M.L.B., B.L., P.E.F., M.E., H.L., V.R.); the Service de Médecine Interne, Groupe Hospitalier PitiéSalpêtrière, Paris (J.-C.P.); the Laboratoire de Bactériologie, Hôpital Louis Pradel, Lyons, France (F.V.); the Service de Médecine Interne, Centre Hospitalier Lyon Sud, Lyons, France (D.V.-D.); and the Department of Medicine, University of Alberta, Edmonton, Canada (T.J.M.).
Address reprint requests to Dr. Raoult at the Unité des Rickettsies, Université de la Méditerranée, Faculté de Médecine, CNRS UPRESA 6020, 27 Blvd. Jean Moulin, 13385 Marseilles, France, or at didier.raoult{at}medecine.univ-mrs.fr.
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