title: A highly efficient Ziehl-Neelsen stain: identifying de novo intracellular Mycobacterium tuberculosis and improving detection of extracellular M. tuberculosis in cerebrospinal fluid.
abstract: Tuberculous meningitis leads to a devastating outcome, and early diagnosis and rapid chemotherapy are vital to reduce morbidity and mortality. Since Mycobacterium tuberculosis is a kind of cytozoic pathogen and its numbers are very few in cerebrospinal fluid, detecting M. tuberculosis in cerebrospinal fluid from tuberculous meningitis patients is still a challenge for clinicians. Ziehl-Neelsen stain, the current feasible microbiological method for the diagnosis of tuberculosis, often needs a large amount of cerebrospinal fluid specimen but shows a low detection rate of M. tuberculosis. Here, we developed a modified Ziehl-Neelsen stain, involving cytospin slides with Triton processing, in which only 0.5 ml of cerebrospinal fluid specimens was required. This method not only improved the detection rate of extracellular M. tuberculosis significantly but also identified intracellular M. tuberculosis in the neutrophils, monocytes, and lymphocytes clearly. Thus, our modified method is more effective and sensitive than the conventional Ziehl-Neelsen stain, providing clinicians a convenient yet powerful tool for rapidly diagnosing tuberculous meningitis.
authors.affiliation: Department of Neurology, Xijing Hospital, the Fourth Military Medical University, Xi’an, Shaanxi 710032, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Chen
authors.forename: Ming
authors.initials: M
authors.lastname: Shi
authors.forename: Guo-Dong
authors.initials: GD
authors.lastname: Feng
authors.forename: Jia-Yun
authors.initials: JY
authors.lastname: Liu
authors.forename: Bing-Ju
authors.initials: BJ
authors.lastname: Wang
authors.forename: Xiao-Dan
authors.initials: XD
authors.lastname: Shi
authors.forename: Lei
authors.initials: L
authors.lastname: Ma
authors.forename: Xue-Dong
authors.initials: XD
authors.lastname: Liu
authors.forename: Yi-Ning
authors.initials: YN
authors.lastname: Yang
authors.forename: Wen
authors.initials: W
authors.lastname: Dai
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Liu
authors.forename: Ying
authors.initials: Y
authors.lastname: He
authors.forename: Jin-Ge
authors.initials: JG
authors.lastname: Li
authors.forename: Xiao-Ke
authors.initials: XK
authors.lastname: Hao
authors.forename: Gang
authors.initials: G
authors.lastname: Zhao
mesh_terms: D004396:Coloring Agents / Q000737:chemistry*; D006801:Humans; D008214:Lymphocytes / Q000382:microbiology; D009169:Mycobacterium tuberculosis; D009504:Neutrophils / Q000382:microbiology; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods*; D014390:Tuberculosis, Meningeal / Q000134:cerebrospinal fluid* / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D004396:Coloring Agents