Culture and immunological detection of Tropheryma whippelii from the duodenum of a patient with Whipple disease

Context Culture of Tropheryma whippelii has been established only once, in human fibroblast cell lines from a heart valve inoculum. Molecular-based di agnostic techniques, although highly sensitive, may be less specific. New d iagnostic tools involving isolation of bacteria from contaminated intestina l biopsies and immunohistological detection need to be developed. Objective To describe a novel method for detection and culture of T whippel ii strains. Design, Setting, and Subjects Laboratory analysis of duodenal biopsy specim ens from a patient with typical relapsing Whipple disease with intestinal i nvolvement, performed Marseille, France, in March 2000, Biopsy specimens we re decontaminated with antimicrobial agents and inoculated onto cell cultur es. Mouse anti-T whippelii polyclonal antibodies were used to detect T whip pelii in fixed specimens taken from the patient before and after relapse, c ompared with specimens from 10 controls. The genotype of the isolate was de termined by amplification and sequencing of 2 DNA fragments (ITS and 23S rR NA). Main Outcome Measure Isolation and genotyping of a new strain(s) of T whipp elii from the case patient's biopsy specimens. Results A strain was grown from the case patient's intestinal specimen that has a genotype different from the first strain isolated, During 2 episodes of Whipple disease, T whippelii bacteria were detected by immunochemistry in the patient's duodenal biopsy specimens, but not in controls. Conclusions A second strain of T whippelii has been isolated and a protocol for isolation from the intestine has been proven to be efficient. Immunode tection of T whippelii in intestinal biopsy specimens may provide a useful tool for the diagnosis and follow-up of patients with Whipple disease. Both techniques need further evaluation and confirmation.