• Guo-dong Feng, Ming Shi, Lei Ma, Ping Chen, Bing-ju Wang, Min Zhang, Xiao-lin Chang, Xiu-chu Su, Yi-ning Yang, Xin-hong Fan, Wen Dai, Ting-ting Liu, Ying He, Ting Bian, Li-xin Duan, Jin-ge Li, Xiao-ke Hao, Jia-yun Liu, Xin Xue, Yun-zhang Song, Hai-qin Wu, Guo-qiang Niu, Li Zhang, Cui-juan Han, Hong Lin, Zhi-hui Lin, Jian-jun Liu, Qian Jian, Jin-she Zhang, Ye Tian, Bai-yu Zhou, Jing Wang, Chang-hu Xue, Xiao-fang Han, Jian-feng Wang, Shou-lian Wang, Guy E Thwaites, and Gang Zhao.
• 1 Department of Neurology.
• Am. J. Respir. Crit. Care Med. 2014 Feb 15; 189 (4): 475481475-81.

RationaleEarly diagnosis and treatment of tuberculous meningitis saves lives, but current laboratory diagnostic tests lack sensitivity.ObjectivesWe investigated whether the detection of intracellular bacteria by a modified Ziehl-Neelsen stain and early secretory antigen target (ESAT)-6 in cerebrospinal fluid leukocytes improves tuberculous meningitis diagnosis.MethodsCerebrospinal fluid specimens from patients with suspected tuberculous meningitis were stained by conventional Ziehl-Neelsen stain, a modified Ziehl-Neelsen stain involving cytospin slides with Triton processing, and an ESAT-6 immunocytochemical stain. Acid-fast bacteria and ESAT-6-expressing leukocytes were detected by microscopy. All tests were performed prospectively in a central laboratory by experienced technicians masked to the patients' final diagnosis.Measurements And Main ResultsTwo hundred and eighty patients with suspected tuberculous meningitis were enrolled. Thirty-seven had Mycobacterium tuberculosis cultured from cerebrospinal fluid; 40 had a microbiologically confirmed alternative diagnosis; the rest had probable or possible tuberculous meningitis according to published criteria. Against a clinical diagnostic gold standard the sensitivity of conventional Ziehl-Neelsen stain was 3.3% (95% confidence interval, 1.6-6.7%), compared with 82.9% (95% confidence interval, 77.4-87.3%) for modified Ziehl-Neelsen stain and 75.1% (95% confidence interval, 68.8-80.6%) for ESAT-6 immunostain. Intracellular bacteria were seen in 87.8% of the slides positive by the modified Ziehl-Neelsen stain. The specificity of modified Ziehl-Neelsen and ESAT-6 stain was 85.0% (95% confidence interval, 69.4-93.8%) and 90.0% (95% confidence interval, 75.4-96.7%), respectively.ConclusionsEnhanced bacterial detection by simple modification of the Ziehl-Neelsen stain and an ESAT-6 intracellular stain improve the laboratory diagnosis of tuberculous meningitis.

33 keywords...

hide…