Aix Marseille Université, URMITE, UMR 6236, CNRS 7278, IRD 198, INSERM 1095, IFR 48, Méditerranée Infection, Faculté de Médecine , Marseille , France.
Front Public Health. 2015 Nov 11;3:254. doi: 10.3389/fpubh.2015.00254. eCollection 2015.
In Africa, relapsing fevers caused by ectoparasite-borne Borrelia species are transmitted by ticks, with the exception of Borrelia recurrentis, which is a louse-borne spirochete. These tropical diseases are responsible for mild to deadly spirochetemia. Cultured Borrelia crocidurae, Borrelia duttonii, and Borrelia hispanica circulate alongside at least six species that have not yet been cultured in vectors. Direct diagnosis is hindered by the use of non-specific laboratory tools. Indeed, microscopic observation of Borrelia spirochaeta in smears of peripheral blood taken from febrile patients lacks sensitivity and specificity. Although best visualized using dark-field microscopy, the organisms can also be detected using Wright-Giemsa or acridine orange stains. PCR-based detection of specific sequences in total DNA extracted from a specimen can be used to discriminate different relapsing fever Borreliae. In our laboratory, we developed a multiplex real-time PCR assay for the specific detection of B. duttonii/recurrentis and B. crocidurae: multispacer sequence typing accurately identified cultured relapsing fever borreliae and revealed diversity among them. Other molecular typing techniques, such as multilocus sequence analysis of tick-borne relapsing fever borreliae, showed the potential risk of human infection in Africa. Recent efforts to culture and sequence relapsing fever borreliae have provided new information for reassessment of the diversity of these bacteria. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been reported as a means of identifying cultured borreliae and of identifying both vectors and vectorized pathogens such as detecting relapsing fever borreliae directly in ticks. The lack of a rapid diagnosis test restricts the management of such diseases. We produced monoclonal antibodies against B. crocidurae in order to develop cheap assays for the rapid detection of relapsing fever borreliae. In this paper, we review point-of-care diagnosis and confirmatory methods.
在非洲,由节肢动物传播的螺旋体 Borrelia 物种引起的回归热通过蜱传播,Borrelia recurrentis 除外,它是一种虱传螺旋体。这些热带疾病可导致轻度至致命性螺旋体血症。培养的Borrelia crocidurae、Borrelia duttonii 和 Borrelia hispanica 与至少六种尚未在媒介中培养的物种一起循环。由于使用非特异性实验室工具,直接诊断受到阻碍。实际上,从发热患者的外周血涂片上观察 Borrelia spirochaeta 的微观观察缺乏敏感性和特异性。尽管使用暗场显微镜观察效果最佳,但也可以使用 Wright-Giemsa 或吖啶橙染色来检测这些生物体。从标本中提取的总 DNA 中基于 PCR 的特定序列检测可用于区分不同的回归热螺旋体 Borreliae。在我们的实验室中,我们开发了一种用于特异性检测 B. duttonii/recurrentis 和 B. crocidurae 的多重实时 PCR 检测方法:多间隔序列分型准确地识别了培养的回归热螺旋体 Borreliae,并揭示了它们之间的多样性。其他分子分型技术,如蜱传回归热螺旋体 Borreliae 的多位点序列分析,显示了非洲人类感染的潜在风险。最近对回归热螺旋体 Borreliae 的培养和测序工作提供了重新评估这些细菌多样性的新信息。最近,基质辅助激光解吸/电离飞行时间质谱被报道为一种识别培养的 Borreliae 以及识别媒介和媒介化病原体的方法,例如直接在蜱中检测回归热螺旋体 Borreliae。缺乏快速诊断测试限制了这些疾病的管理。我们针对 B. crocidurae 产生了单克隆抗体,以便开发用于快速检测回归热螺旋体 Borreliae 的廉价检测方法。本文综述了即时诊断和确认方法。