Jafari Claudia, Ernst Martin, Kalsdorf Barbara, Greinert Ulf, Diel Roland, Kirsten Detlef, Marienfeld Kathleen, Lalvani Ajit, Lange Christoph
Division of Clinical Infectious Diseases, Research Center Borstel, Parkallee 35, Borstel, Germany.
Am J Respir Crit Care Med. 2006 Nov 1;174(9):1048-54. doi: 10.1164/rccm.200604-465OC. Epub 2006 Jul 20.
In a large proportion of patients with active pulmonary tuberculosis (pTB), acid-fast bacilli smear results for sputum and bronchial secretions are negative. Detectable growth of Mycobacterium tuberculosis (MTB) in cultures takes several weeks and MTB-specific DNA amplification results on sputum and bronchial secretions are variable in these patients.
We investigated whether a rapid diagnosis of pTB can be established by enumeration of MTB-specific mononuclear cells from bronchoalveolar lavage (BAL) fluid in routine clinical practice.
Patients presenting to a tertiary hospital with medical histories and pulmonary infiltrates compatible with tuberculosis, and negative acid-fast bacilli smear results (three) from sputum, were prospectively enrolled in this study. An MTB-specific enzyme-linked immunospot assay (ELISPOT [T-SPOT.TB; Oxford Immunotec, Abingdon, UK]) with early antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10) peptides was performed on peripheral blood mononuclear cells (PBMCs) and mononuclear cells from the BAL fluid (BALMCs).
Of 37 patients, 12 were found to have smear-negative pTB and 25 were found to have an alternative diagnosis. Patients with tuberculosis had a median number of 17 ESAT-6-specific cells and 24.5 CFP-10-specific cells per 200,000 PBMCs and 37.5 ESAT-6-specific cells and 49.5 CFP-10-specific cells per 200,000 cells in the BAL fluid. Control patients had a median of 1 ESAT-6-specific cell and 1 CFP-10-specific cell per 200,000 PBMCs and no ESAT-6- and CFP-10-specific cells per 200,000 cells in the BAL fluid (p < 0.0001). All patients with TB but none of the control subjects had more than 5 spot-forming cells per 200,000 BALMCs with either peptide in the BAL fluid ELISPOT.
Smear-negative pulmonary tuberculosis can be diagnosed rapidly by identification of MTB-specific cells in the BAL fluid.
在大部分活动性肺结核（pTB）患者中，痰液和支气管分泌物的抗酸杆菌涂片结果为阴性。结核分枝杆菌（MTB）培养中可检测到的生长需要数周时间，并且这些患者痰液和支气管分泌物的MTB特异性DNA扩增结果存在差异。
我们调查了在常规临床实践中，能否通过对支气管肺泡灌洗（BAL）液中的MTB特异性单核细胞进行计数来快速诊断pTB。
前瞻性纳入一家三级医院中具有与结核病相符的病史和肺部浸润且痰液抗酸杆菌涂片结果为阴性（三次）的患者。对外周血单核细胞（PBMC）和BAL液中的单核细胞（BALMC）进行针对早期抗原靶点-6（ESAT-6）和培养滤液蛋白-10（CFP-10）肽段的MTB特异性酶联免疫斑点试验（ELISPOT [T-SPOT.TB；牛津免疫技术公司，英国阿宾顿]）。
37例患者中，12例被发现患有涂片阴性的pTB，25例被发现患有其他诊断疾病。结核病患者每200,000个PBMC中ESAT-6特异性细胞的中位数为17个，CFP-10特异性细胞的中位数为24.5个；BAL液中每200,000个细胞中ESAT-6特异性细胞的中位数为37.5个，CFP-10特异性细胞的中位数为49.5个。对照患者每200,000个PBMC中ESAT-6特异性细胞的中位数为1个，CFP-10特异性细胞的中位数为1个；BAL液中每200,000个细胞中无ESAT-6和CFP-10特异性细胞（p < 0.0001）。在BAL液ELISPOT中，所有结核病患者但无对照受试者每200,000个BALMC中任何一种肽段的斑点形成细胞超过5个。
通过识别BAL液中的MTB特异性细胞可快速诊断涂片阴性的肺结核。
