Asmar Shady, Drancourt Michel
Faculté de Médecine, URMITE, UM63, Centre National de la Recherche Scientifique 7278, IRD 198, Institut National de la Santé et de la Recherche Médicale 1095, Aix Marseille Université Marseille, France.
Front Microbiol. 2015 Nov 3;6:1184. doi: 10.3389/fmicb.2015.01184. eCollection 2015.
Culturing Mycobacterium tuberculosis remains the gold standard for the laboratory diagnosis of pulmonary tuberculosis, with 9 million new cases and 1.5 million deaths mainly in developing countries. Reviewing data reported over 20 years yields a state-of-the-art procedure for the routine culture of M. tuberculosis in both developed and developing countries. Useful specimens include sputum, induced sputum, and stools collected in quaternary ammonium preservative-containing sterile cans. The usefulness of other non-invasive specimens remains to be evaluated. Specimens can be collected in a diagnosis kit also containing sampling materials, instructions, laboratory requests, and informed consent. Automated direct LED fluorescence microscopy after auramine staining precedes inoculation of an egg-lecithin-containing culture solid medium under microaerophilic atmosphere, inverted microscope reading or scanning video-imaging detection of colonies and colonies identification by recent molecular methods. This procedure should result in a diagnosis of pulmonary tuberculosis as fast as 5 days. It may be implemented in both developed and developing countries with automated steps replaceable by manual steps depending on local resources.
培养结核分枝杆菌仍然是肺结核实验室诊断的金标准,肺结核每年有900万新发病例,主要在发展中国家导致150万人死亡。回顾20多年来报告的数据,得出了一套发达国家和发展中国家常规培养结核分枝杆菌的先进方法。有用的标本包括痰液、诱导痰以及收集于含季铵防腐剂无菌罐中的粪便。其他非侵入性标本的有用性仍有待评估。标本可收集于诊断试剂盒中,该试剂盒还包含采样材料、说明、实验室申请单及知情同意书。金胺染色后进行自动直接LED荧光显微镜检查,然后在微需氧环境下接种含卵黄卵磷脂的培养固体培养基,通过倒置显微镜读数或扫描视频成像检测菌落,并采用最新分子方法进行菌落鉴定。该方法应能在5天内快速诊断出肺结核。在发达国家和发展中国家均可实施,根据当地资源情况,自动化步骤可用手工步骤替代。