Raoult D, La Scola B, Lecocq P, Lepidi H, Fournier P E
Unité des Rickettsies, Faculté de Médecine, Université des la Méditerranée/CNRS, 27 Boulevard Jean Moulin, UPRESA 6020, 13385 Marseille CEDEX 05, France.
JAMA. 2001 Feb 28;285(8):1039-43. doi: 10.1001/jama.285.8.1039.
Culture of Tropheryma whippelii has been established only once, in human fibroblast cell lines from a heart valve inoculum. Molecular-based diagnostic techniques, although highly sensitive, may be less specific. New diagnostic tools involving isolation of bacteria from contaminated intestinal biopsies and immunohistological detection need to be developed.
To describe a novel method for detection and culture of T whippelii strains.
DESIGN, SETTING, AND SUBJECTS: Laboratory analysis of duodenal biopsy specimens from a patient with typical relapsing Whipple disease with intestinal involvement, performed Marseille, France, in March 2000. Biopsy specimens were decontaminated with antimicrobial agents and inoculated onto cell cultures. Mouse anti-T whippelii polyclonal antibodies were used to detect T whippelii in fixed specimens taken from the patient before and after relapse, compared with specimens from 10 controls. The genotype of the isolate was determined by amplification and sequencing of 2 DNA fragments (ITS and 23S rRNA).
Isolation and genotyping of a new strain(s) of T whippelii from the case patient's biopsy specimens.
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.
A second strain of T whippelii has been isolated and a protocol for isolation from the intestine has been proven to be efficient. Immunodetection 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.
惠普尔嗜组织菌仅在一次实验中成功培养，所用接种物是取自心脏瓣膜的人类成纤维细胞系。基于分子的诊断技术虽然高度敏感，但特异性可能较差。需要开发新的诊断工具，包括从受污染的肠道活检标本中分离细菌以及进行免疫组织学检测。
描述一种检测和培养惠普尔嗜组织菌菌株的新方法。
设计、地点和研究对象：2000年3月在法国马赛对一名患有典型复发性惠普尔病且累及肠道的患者的十二指肠活检标本进行实验室分析。活检标本用抗菌剂进行净化处理后接种到细胞培养物上。使用小鼠抗惠普尔嗜组织菌多克隆抗体检测该患者复发前后的固定标本中的惠普尔嗜组织菌，并与10名对照者的标本进行比较。通过对两个DNA片段（ITS和23S rRNA）进行扩增和测序来确定分离株的基因型。
从病例患者的活检标本中分离出新的惠普尔嗜组织菌菌株并进行基因分型。
从病例患者的肠道标本中培养出一种菌株，其基因型与首次分离出的菌株不同。在惠普尔病的两次发作期间，通过免疫化学方法在患者的十二指肠活检标本中检测到了惠普尔嗜组织菌，但在对照者标本中未检测到。
已分离出第二株惠普尔嗜组织菌，并且已证明从肠道分离该菌的方案是有效的。在肠道活检标本中对惠普尔嗜组织菌进行免疫检测可能为惠普尔病患者的诊断和随访提供一种有用的工具。这两种技术都需要进一步评估和确认。
