检测脑脊液中CFP10和ESAT-6对结核性脑膜炎诊断意义的研究
|
摘要
目的:改进斑点酶联免疫吸附试验(dot enzyme linked immunosorbant assay, Dot-ELISA),采用真空泵抽吸法浓缩检测样本,以期提高此方法的敏感性;通过实验分析此方法改进前后的优劣,并确定试验程序中最适工作条件及方法的重复性和稳定性。应用改进的Dot-ELISA检测脑脊液(cerebrospinal fluid,CSF)中结核分枝杆菌(mycobacterium tuberculosis,MTB)特异性抗原培养滤液蛋白10(culture filtrate protein 10,CFP10)和6000早期分泌性抗原靶(6kDa early secretory antigenic target,ESAT-6),对比其他辅助检查及同类研究,评价其对结核性脑膜炎(tuberculous meningitis,TBM)的诊断价值。
方法:
1、用改进前和改进后的Dot-ELISA检测同一批临床确诊的结核性胸腔积液25例和非结核性胸腔积液31例,统计分析方法改进前后有无差异性;改变实验程序中的工作条件,利用对比实验确定最适工作条件,并评价改进后此方法的重复性和稳定性。2、收集58例临床诊为TBM患者的CSF,53例非TBM患者的CSF,应用改进后的Dot-ELISA分别检测CSF中CFP10和ESAT-6,同时对入选的患者和采集的CSF作相关的病史统计和辅助检查,并对结果进行统计分析。
结果:
1、改进前Dot-ELISA检测的敏感性为76.0%,改进后的敏感性为92.0%,用最适工作条件检测的敏感性为96.0%,改进后和用最适工作条件与改进前比较存在统计学差异(P<0.05);改进前特异性为93.5%,改进后和用最适条件时特异性皆为90.3%,改进前后特异性比较无统计学差异(P>0.05),验证其特异性的阻断实验结果为阳性;重复性、稳定性试验结果表明,所建立的Dot-ELISA检测结果的可重复率为100%。2、检测CSF中CFP10抗原的敏感性为93.1%,特异性为92.5%;检测ESAT-6抗原的敏感性为91.4%,特异性为94.3%;敏感性和特异性均普遍高于针对MTB菌体或菌体物质进行的检测及同类研究,如抗酸染色、结核杆菌培养、PCR等。
结论:
1、所建立的Dot-ELISA是一种有较高敏感性和特异性且重复性和稳定性较好的检测方法,能用于结核特异性抗原的检测和结核性胸腔积液的辅助诊断。2、应用改进的Dot-ELISA检测CSF中CFP10和ESAT-6对诊断TBM有很高的敏感性和特异性,明显优于针对MTB菌体或菌体物质进行的检测和同类研究,目前其可作为优于其它辅助检查的一项新的检查手段用于临床TBM的诊断。3、本研究为MTB特异性抗原用于结核病的诊断奠定了基础。
Objection: Our study improves dot enzyme linked immunosorbant assay (Dot-ELISA), using vacuum pump to aspirate the water in specimens to concentrate them, in order to increase the sensitivity of the method. Use experiments to analyze advantages and disadvantages of the improved method, and to determine the best working condition and the repeatability and the stability of the developed method. Using the improved Dot-ELISA method to detect culture filtrate protein 10 (CFP-10) and 6000 early secretory antigenic target (ESAT-6), two small protein antigen secreted by M.tuberculosis (MTB), in cerebrospinal fluid (CSF). And compare them with other examination and studies in order to judge and analyse their value in diagnosing tuberculous meningitis (TBM).
Methods: We detected the same batch of pleural effusion using the old method and the developed method respectively, which include 25 specimens of tuberculous pleural effusion and 31 non-tuberculous and analyzed the difference of the two methods. Determine the repeatability and stability as well as the best working condition by changing the working condition. 111 specimens of cerebrospinal fluid were collected, in which 58 specimens were clinically diagnosed as tuberculous meningitis and 53 as non-tuberlous. CFP10 and ESAT-6 were detected in the cerebrospinal fluid using Dot ELISA method and the results were analyzed.
Results: The sensibility of the old method was 76.0%, the improvement method was 92.0%. The sensibility was 96.0% when the best working condition used in the method. There existed statistical difference between the old method and the improvement method as well as using the best working condition (P<0.05). The specificity of the old method was 93.5%. The specificity of improvement method and it in the best working condition was 90.3%. But the specificity doesn’t have statistical difference. Blocking experiment which can verify its specificity was positive.The repeatability and stability were good, particularly the repetition rate of the positive results was 100%. The sensitivities of detecting CFP10 and ESAT-6 antigen were 93.1% and 91.4% respectively, and the specificities were 92.5% and 94.3% respectively. The sensitivities and specificities are generally higher compared with those detecting M.tuberculosis or materials of M.tuberculosis, for example, acid-fast staining or mycobacterium tuberculosis culture or PCR.
Conclusion: It shows that the developed Dot-ELISA method is sensitive, specific, stable, and can be used to detect specific antigen of mycobacterium tuberculosis and diagnosis of the tuberculosis.Using Dot-ELISA method to detect CFP10 and ESAT-6 in cerebrospinal fluid to diagnose tuberculous meningitis has a high detection rate. The sensitivities and specificities are generally higher compared with those detecting M.tuberculosis or materials of M.tuberculosis. It can be used in diagnosing tuberculous meningitis as a new method which is better than other methods. Our study provided the evidence for the specific antigen of M.tuberculosis used in diagnosing tuberculosis.
方法:
1、用改进前和改进后的Dot-ELISA检测同一批临床确诊的结核性胸腔积液25例和非结核性胸腔积液31例,统计分析方法改进前后有无差异性;改变实验程序中的工作条件,利用对比实验确定最适工作条件,并评价改进后此方法的重复性和稳定性。2、收集58例临床诊为TBM患者的CSF,53例非TBM患者的CSF,应用改进后的Dot-ELISA分别检测CSF中CFP10和ESAT-6,同时对入选的患者和采集的CSF作相关的病史统计和辅助检查,并对结果进行统计分析。
结果:
1、改进前Dot-ELISA检测的敏感性为76.0%,改进后的敏感性为92.0%,用最适工作条件检测的敏感性为96.0%,改进后和用最适工作条件与改进前比较存在统计学差异(P<0.05);改进前特异性为93.5%,改进后和用最适条件时特异性皆为90.3%,改进前后特异性比较无统计学差异(P>0.05),验证其特异性的阻断实验结果为阳性;重复性、稳定性试验结果表明,所建立的Dot-ELISA检测结果的可重复率为100%。2、检测CSF中CFP10抗原的敏感性为93.1%,特异性为92.5%;检测ESAT-6抗原的敏感性为91.4%,特异性为94.3%;敏感性和特异性均普遍高于针对MTB菌体或菌体物质进行的检测及同类研究,如抗酸染色、结核杆菌培养、PCR等。
结论:
1、所建立的Dot-ELISA是一种有较高敏感性和特异性且重复性和稳定性较好的检测方法,能用于结核特异性抗原的检测和结核性胸腔积液的辅助诊断。2、应用改进的Dot-ELISA检测CSF中CFP10和ESAT-6对诊断TBM有很高的敏感性和特异性,明显优于针对MTB菌体或菌体物质进行的检测和同类研究,目前其可作为优于其它辅助检查的一项新的检查手段用于临床TBM的诊断。3、本研究为MTB特异性抗原用于结核病的诊断奠定了基础。
Objection: Our study improves dot enzyme linked immunosorbant assay (Dot-ELISA), using vacuum pump to aspirate the water in specimens to concentrate them, in order to increase the sensitivity of the method. Use experiments to analyze advantages and disadvantages of the improved method, and to determine the best working condition and the repeatability and the stability of the developed method. Using the improved Dot-ELISA method to detect culture filtrate protein 10 (CFP-10) and 6000 early secretory antigenic target (ESAT-6), two small protein antigen secreted by M.tuberculosis (MTB), in cerebrospinal fluid (CSF). And compare them with other examination and studies in order to judge and analyse their value in diagnosing tuberculous meningitis (TBM).
Methods: We detected the same batch of pleural effusion using the old method and the developed method respectively, which include 25 specimens of tuberculous pleural effusion and 31 non-tuberculous and analyzed the difference of the two methods. Determine the repeatability and stability as well as the best working condition by changing the working condition. 111 specimens of cerebrospinal fluid were collected, in which 58 specimens were clinically diagnosed as tuberculous meningitis and 53 as non-tuberlous. CFP10 and ESAT-6 were detected in the cerebrospinal fluid using Dot ELISA method and the results were analyzed.
Results: The sensibility of the old method was 76.0%, the improvement method was 92.0%. The sensibility was 96.0% when the best working condition used in the method. There existed statistical difference between the old method and the improvement method as well as using the best working condition (P<0.05). The specificity of the old method was 93.5%. The specificity of improvement method and it in the best working condition was 90.3%. But the specificity doesn’t have statistical difference. Blocking experiment which can verify its specificity was positive.The repeatability and stability were good, particularly the repetition rate of the positive results was 100%. The sensitivities of detecting CFP10 and ESAT-6 antigen were 93.1% and 91.4% respectively, and the specificities were 92.5% and 94.3% respectively. The sensitivities and specificities are generally higher compared with those detecting M.tuberculosis or materials of M.tuberculosis, for example, acid-fast staining or mycobacterium tuberculosis culture or PCR.
Conclusion: It shows that the developed Dot-ELISA method is sensitive, specific, stable, and can be used to detect specific antigen of mycobacterium tuberculosis and diagnosis of the tuberculosis.Using Dot-ELISA method to detect CFP10 and ESAT-6 in cerebrospinal fluid to diagnose tuberculous meningitis has a high detection rate. The sensitivities and specificities are generally higher compared with those detecting M.tuberculosis or materials of M.tuberculosis. It can be used in diagnosing tuberculous meningitis as a new method which is better than other methods. Our study provided the evidence for the specific antigen of M.tuberculosis used in diagnosing tuberculosis.
引文
[1] Thwaites GE, Chau TT, Stepniewska K, et al. Diagnosis of adult tuberculous meningitis by use of clinical and laboratory features. Lancet, 2002, 360(9342): 1287-1292.
[2] Thwaites GE, Nguyen DB,Nquyen HD,et al. Dexamethasone for the treatment of tuberculous meningitis in adolescents and adults. N Engl J Med, 2004, 351(17): 1741-1751.
[3] Vinnard C, Macgregor RR.Tuberculous meningitis in HIV-infected individuals. Curr HIV/AIDS Rep, 2009 ,6(3):139-45.
[4] American Thoracic Society and Centers for Disease Control and Prevention.2000. Targeted tuberculin testing and treatment of latent tuberculosis infection. Am. J. Respir. Crit. Care Med. 161:S221–S247.
[5] Kulkarni SP, Jaleel MA, Kadival GV. Evaluation of an in house developed PCR for the diagnosis of tuberculous meningitis in Indian children. J Med Microbiol, 2005, 54(Pt4): 369-373.
[6] Haldar S, Sharma N, Gupta VK, et al. Efficient diagnosis of tuberculous meningitis by detection of Mycobacterium tuberculosis DNA in cerebrospinal fluid filtrates using PCR.J Med Microbiol, 2009,58(Pt 5):616-624.
[7] Kashyap RS,Kainthla RP,Biswas SK,et al.Rapid diagnosis of tuberculous meningitis using the simple Dot ELISA method.Med Sci Monit, 2003, 9(11): MT123-126.
[8] Kashyap RS, Dobos KM, Belisle JT,et al.Demonstration of components of antigen 85 complex in cerebrospinal fluid of tuberculous meningitis patients.Clin Diagn Lab Immunol ,2005,12 (6) :752-758.
[9] Quan C, Lu CZ, Qiao J,et al.Comparative evaluation of early diagnosis of tuberculous meningitis by different assays.J Clin Microbiol, 2006, 44(9): 3160-3166.
[10] Losi, M, Bossink A, Codecasa L, et al. Use of a T-cell interferon-gamma release assay for the diagnosis of tuberculous pleurisy. Eur Respir J, 2007, 30(6): 1173-1179.
[11] Mathai A,Radhakrishnan VV,Sarada C,et al.Detection of heat stable mycobacterial antigen in cerebrospinal fluid by Dot-Immunobinding assay. Neurol India, 2003 , 51(1):52-54.
[12] Demkow U,Filewska M,Bialas B,et al.Antimycobacterial antibody level in pleural pericardial and cerebrospinal fluid of patients with tuberculosis.Pneumonol Alergol Pol, 2004, 72(3-4):105-110.
[13] Kashyap RS,Kainithla RP,Satpute RM,et al. Demonstration of IgG antibodies to 30 Kd protein antigen in CSF for diagnosis of tuberculous meningitis by antibody capturing ELISA. Neurol India, 2004,52 (3) :359-362.
[14] Quan C, Lu CZ, Qiao J,et al. Comparative evaluation of early diagnosis of tuberculous meningitis by different assays. J Clin Microbiol, 2006, 44(9): 3160-3166.
[15] Luca MC, Petrovici CM, Va?? A, et al.Gamma interferon testing in blood and cerebrospinalfluid--rapid method for the diagnosis of tuberculous meningitis. Rev Med Chir Soc Med Nat Iasi, 2008 ,112(1):108-110.
[16] Thomas MM, Hinks TS, Raghuraman S, et al. Rapid diagnosis of Mycobacterium tuberculosis meningitis by enumeration of cerebrospinal fluid antigen-specific T-cells. Int J Tuberc Lung Dis, 2008 ,12(6): 651-657.
[17] Murakami S, Takeno M, Oka H,et al. Diagnosis of Tuberculous Meningitis Due to Detection of ESAT-6-Specific Gamma Interferon Production in Cerebrospinal Fluid Enzyme-Linked Immunospot Assay.Clinical and Vaccine Immunol,2008,15(5) 897-899.
[18] Srivastava KL, Bansal M, Gupta S,et al.Diagnosis of tuberculous meningitis by detection of antigen and antibodies in CSF and sera. Indian Pediatr. 1998, 35(9): 841-50.
[19] Restrepo BI, Pino PA, Volcy M, et al.Interpretation of mycobacterial antibodies in the cerebrospinal fluid of adults with tuberculous meningitis.Trop Med Int Health, 2008, 13(5):653-658.
[20] Venkatesh K, Parija SC, Mahadevan S,et al.Reverse passive haemagglutination (RPHA) test for detection of mycobacterial antigen in the cerebrospinal fluid for diagnosis of tubercular meningitis. Indian J Tuberc,2007,54(1):41-48.
[21] Sumi MG, Mathai A, Reuben S, et al.Immunocytochemical method for early laboratory diagnosis of tuberculous meningitis. Clin Diagn Lab Immunol, 2002, 9(2):344-347.
[22] Renshaw PS,Panagiotidou P,Whelan A,et al.Conclusive evidence that the major T-cell antigens of the Mycobacterium tuberculosis complex ESAT-6 and CFP-10 form a tight,1:1 complex and characterization of the structural properties of ESAT-6, CFP-10,and the ESAT-6*CFP-10 complex.Implications for pathogenesis and virulence. J Biol Chem, 2002, 277:21598–21603.
[23] Converse, S E, and JS. A protein secretion pathway critical for Mycobacterium tuberculosis virulence is conserved and functional in Mycobacterium smegmatis. J Bacteriol, 2005, 187:1238–1245.
[24] Lewis KN, Liao R, Guinn K M,et al. Deletion of RD1 from Mycobacterium tuberculosis mimics bacille Calmette-Guerin attenuation. J Infect Dis, 2003, 187:117–123.
[25] Amoudy HA, Al-Turab MB, Mustafa AS. Identification of transcriptionally active open reading frames within the RD1 genomic segment of Mycobacterium tuberculosis. Med Princ Pract, 2006, 15:137–144.
[26] van Pinxteren LA, Ravn P, Agger EM, et al. Diagnosis of tuberculosis based on the two specific antigens ESAT-6 and CFP10. Clin Diagn Lab Immunol, 2000, 7(2): 155-160.
[27] Ahuja GK, Mohan KK, Prasad K, Behari M.Diagnostic criteria for tuberculous meningitis and their validation. Tuber Lung Dis, 1994, 75(2):149-152.
[28] Seth R, Sharma U.Diagnostic criteria for Tuberculous Meningitis. Indian J Pediatr, 2002, 69(4):299-303.
[29] Miceli, I., I. N. de Kantor, et al.Evaluation of the effectiveness of BCG vaccination using the case-control method in Buenos Aires, Argentina. Int. J. Epidemiol. 1988, 17:629-634.
[30] Cicero R, Olivera H, Hernández-Solis A,et al.Frequency of Mycobacterium bovis as an etiologic agent in extrapulmonary tuberculosis in HIV-positive and -negative Mexican patients. Eur J Clin Microbiol Infect Dis, 2009, 28(5):455-460.
[31] Shah, N. P., A. Singhal, et al. Occurrence of overlooked zoonotic tuberculosis: detection of Mycobacterium bovis in human cerebrospinal fluid. J. Clin. Microbiol, 2006, 44:1352-1358.
[32] Ritacco V, de Kantor IN.Simultaneous detection of Mycobacterium bovis and Mycobacterium tuberculosis in human cerebrospinal fluid. J Clin Microbiol. 2007, 45(2): 684.
[33] Thwaites G,Caws M,Chau TT,et al.Relationship between Mycobacterium tuberculosis genotype and the clinical phenotype of pulmonary and meningeal tuberculosis. J Clin Microbiol, 2008, 46(4):1363-1368.
[34]王维治,郭玉璞,宿英英.神经病学.人民卫生出版社,2006,1 :670-675.
[35] Flesch I, Gefferth C.X-ray examination of the ventricular system of children suffering from tuberculous meningitis. Paediatr Danub, 1949, 5(3-4):158-168.
[36] Yarami? A, Bükte Y, Katar S, et al.Chest computerized tomography scan findings in 74 children with tuberculous meningitis in southeastern Turkey. Turk J Pediatr, 2007, 49(4): 365-369.
[37] Andronikou S, Wieselthaler N, Smith B,et al.Value of early follow-up CT in paediatric tuberculous meningitis. Pediatr Radiol, 2005, 35(11):1092-1099.
[38] van der Merwe DJ, Andronikou S, Van Toorn R, et al. Brainstem ischemic lesions on MRI in children with tuberculous meningitis: with diffusion weighted confirmation. Childs Nerv Syst, 2009, 25(8):949-954.
[39] Pienaar M, Andronikou S, van Toorn R.MRI to demonstrate diagnostic features and complications of TBM not seen with CT. Childs Nerv Syst, 2009, 25(8):941-947.
[40] Chitre PS, Tullu MS, Sawant HV, et al. Co-occurrence of intracerebral tuberculoma with lumbar intrame dullary tuberculoma. J Child Neurol, 2009, 24(5):606-609.
[41] Xiong CH, Liang XF, Wang HQ. A systematic review on the protective efficacy of BCG against children tuberculosis meningitis and millet tuberculosis. Zhongguo Ji Hua Mian Yi, 2009, 15(4):358-362.
[42] Wasay M, Ajmal S, Taqui AM, et al. Impact of Bacille Calmette-Guerin Vaccination on Neuroradiological Manifestations of Pediatric Tuberculous Meningitis. J Child Neurol, 2009, 30(5):60-63.
[43] Dural AT, Johnson PC, Wagner A. The utility of direct acid-fast stain of cerebrospinal fluid for the rapid diagnosis of tuberculous meningitis in patients with and without AIDS. J Gen Intern Med, 1997,12(4):259.
[44] Radhakrishnan VV, Mathai A, Thomas M. Correlation between culture of Mycobacterium tuberculosis and antimycobacterial antibody in lumbar, ventricular and cisternal cerebrospinal fluids of patients with tuberculous meningitis. Indian J Exp Biol, 1991, 29(9): 845-848.
[45] Krishnan VV,Mathai A.Isolation of two antigens from the culture filtrates of Mycobacterium tuberculosis and their applications in the laboratory diagnosis of the tuberculous meningitis.Med Microbiol Immunol,1991,180(2):101-107.
[46] Baveja CP, Gumma V, Jain M, et al.Newer methods over the conventional diagnostic tests for tuberculous meningitis: do they really help? Trop Doct, 2009, 39(1): 18-20.
[47]赵钢.脑脊液细胞学研究的现状和展望.神经损伤与功能重建,2006,1(3):168-170.
[48] Katti MK.Immunodiagnosis of tuberculous meningitis: rapid detection of mycobacterial antigens in cerebrospinal fluid by reverse passive hemagglutination assay and their characterization by Western blotting. FEMS Immunol Med Microliol, 2001, 31(1):59-64 .
[49] Tiwari RP, Garg SK, Bharmal RN,et al.Rapid liposomal agglutination card test for the detection of antigens in patients with active tuberculosis.Int J Tuberc Lung Dis, 2007, 11(10): 1143-1151.
[50] Samuel AM,Kadival GV,Irani S,et al.A sensitive and specific method for diagnosis of tubercular meningitis. Indian J Med Res, 1983, 77:752-757.
[51] Sada E, Ruiz-Palacios G M, López-Vidal Y, et al. Detection of mycobacterial antigens in cerebrospinal ?uid of patients with tuberculous meningitis by enzyme-linked immunosorbent assay.Lancet.1983,2: 651–652.
[52] Krambovitis E, McIllmurray M B, Lock P E, et al. Rapid diagnosis of tuberculous meningitis by latex particle agglutination.Lancet.1984,2: 1229–1231.
[53] Kadival G V, Mazarelo T B, Haparas S D. Sensitivity and specificity of enzyme linked immunosorbent assay in the detection of antigen in tuberculous meningitis cerebrospinal ?uids. J Clin Microbiol. 1986,23: 901–904.
[54] Murakami S, Takeno M, Oka H, et al.Diagnosis of tuberculous meningitis due to detection of ESAT-6-specific gamma interferon production in cerebrospinal fluid enzyme-linked immunospot assay.Clin Vaccine Immunol,2008,15(5):897-899.
[55] Losi, M, Bossink A, Codecasa L, et al. Use of a T-cell interferon-gamma release assay for the diagnosis of tuberculous pleurisy. Eur Respir J, 2007, 30(6): 1173-1179.
[56] Cole ST, Brosch R, Parkhill J, et al. Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence.Nature,1998,393:537–544.
[57] Hart PD, Sutherland I. BCG and vole bacillus vaccines in the prevention of tuberculosis in adolescence andearly adult life. Br Med J, 1977, 2:293–295.
[58] Sorensen AL, Nagai S, Houen G,et al.Purification and characterization of a low-molecular-mass T-cell antigen secreted by Mycobacterium tuberculosis. Infect Immun, 1995,63:1710–1717.
[59] Berthet FX, Rasmussen PB, Rosenkrands I, et al. Mycobacterium tuberculosis operon en-coding ESAT-6 and a novel low-molecular-mass culture filtrate protein (CFP-10). Microbiology, 1998,144:3195–3203.
[60] Brodin P, Rosenkrands I, Andersen P, et al.ESAT-6 proteins: protective antigens and virulence factors? Trends Microbiol, 2004,12:500–508.
[61] Pym AS, Brodin P, Majlessi L,et al.Recombinant BCG exporting ESAT-6 confers enhanced protection against tuberculosis. Nat Med, 2003,9:533–539.
[62] Guinn KM, Hickey MJ, Mathur SK, et al. Individual RD1-region genes are required for export of ESAT-6/CFP-10 and for virulence of Mycobacterium tuberculosis. Mol Microbiol, 2004, 51:359–370.
[63] Gao LY, Guo S, McLaughlin B, et al.A mycobacterial virulence gene cluster extending RD1 is required for cytolysis, bacterial spreading and ESAT-6 secretion. Mol Microbiol, 2004, 53:1677–1693.
[64] Pym AS, Brodin P, Majlessi L, et al. Recombinant BCG exporting ESAT-6 confers enhanced protection against tuberculosis. Nat Med, 2003, 9:533–539.
[65] Lightbody KL, Renshaw PS, Collins ML,et al.Characterisation of complex formation between members of the Mycobacterium tuberculosis complex CFP-10/ESAT-6 protein family:towards an understanding of the rules governing complex formation and thereby functional flexibility. FEMS Microbiol Lett, 2004,238:255–262.
[66] Renshaw PS, Panagiotidou P, Whelan A,et al.Conclusive evidence that the major T-cell antigens of the Mycobacterium tuberculosis complex ESAT-6 and CFP-10 form a tight, 1:1 complex and characterization of the structural properties of ESAT-6, CFP-10,and the ESAT-6*CFP-10 complex. Implications for pathogenesis and virulence. J Biol Chem, 2002,277:21598–21603.
[67] Papps M G, Ajkowski R, Hockmeyer W T. Dot-enzyme-linked immunosorbent assay (dot-ELISA): a micro technique for rapid diagnosis of visceral leishmaniasis. J Immunol Methods, 1983, 64(1-2): 205-214.
[68] Colangeli R, Spencer JS, Bifani P,et al. MTSA-10, the product of the Rv3874 gene of Mycobacterium tuberculosis, elicits tuberculosis-specific, delayed-type hypersensitivity in guinea pigs. Infect Immun, 2000, 68:990–993.
[69] Sahn SA. Pleural thickening, trapped lung, and chronic empyemaas sequelae of tuberculous pleural effusion: don’t sweat the pleuralthickening. Int J Tuberc Lung Dis, 2002, 6: 461-464.
[70] Shenai SC ,Rodrigues, A P Mehta,et al. Evaluation of a new phage Amplification technology for rapid diagnosis of tuberculosis. Indian J. Med. Microbiol, 2002, 20:194–199.
[71] Harboe M, Oettinger T, Wiker HG, et al. Evidence for occurrence of the ESAT-6 protein in Mycobacterium tuberculosis and virulent Mycobacterium bovis and for its absence in Mycobacterium bovis BCG. Infect Immun, 1996, 64:16–22.
[72]全国结核病流行病学抽样调查技术指导组.第四次全国结核病流行病学抽样调查报告.中华结核和呼吸杂志,2002,25(1):6-10.
[1] Thwaites GE, Chau TT,Stepniewska K,et al. Diagnosis of adult tuberculous meningitis by use of clinical and laboratory features[J]. Lancet, 2002,360(9342):1287-1292.
[2] Thwaites GE, Nguyen DB,Nquyen HD,et al. Dexamethasone for the treatment of tuberculous meningitis in adolescents and adults[J]. N Engl J Med, 2004,351(17):1741-1751.
[3] Kulkarni SP, Jaleel MA, Kadival GV. Evaluation of an in-house-developed PCR for the diagnosis of tuberculous meningitis in Indian children[J]. J Med Microbiol, 2005, 54(Pt4):369-373.
[4] Sumi MG, Mathai A, Reuben S, et al.Immunocytochemical method for early laboratory diagnosis of tuberculous meningitis[J]. Clin Diagn Lab Immunol, 2002, 9(2):344-347.
[5] Kashyap RS,Kainthla RP,Biswas SK,et al.Rapid diagnosis of tuberculous meningitis using the simple Dot ELISA method[J].Med Sci Monit,2003,9(11):MT123-126.
[6] Mathai A,Radhakrishnan VV,Sarada C,et al.Detection of heat stable mycobacterial antigen in cerebrospinal fluid by Dot-Immunobinding assay [J].Neurol India,2003 ,51(1):52-54.
[7] Kashyap RS, Dobos KM, Belisle JT,et al.Demonstration of components of antigen 85 complex in cerebrospinal fluid of tuberculous meningitis patients[J].Clin Diagn Lab Immunol ,2005,12 (6) :752-758.
[8] Venkatesh K, Parija SC, Mahadevan S,et al.Reverse passive haemagglutination (RPHA) test for detection of mycobacterial antigen in the cerebrospinal fluid for diagnosis of tubercular meningitis [J].Indian J Tuberc,2007,54(1):41-48.
[9]丁晓旭,薛承岩.脑脊液和血液标本检测阿拉伯糖甘露糖脂G抗体诊断结核性脑膜炎的价值[J ].临床荟萃,2007,22 (4) :284-286.
[10]李永刚,杨昆胜,晋华文,等.脂阿拉伯甘露聚糖诊断结核性脑膜炎的评价[J].中国防痨杂志,2005, 27(5):316-319 .
[11] Demkow U,Filewska M,Bialas B,et al.Antimycobacterial antibody level in pleural , pericardial and cerebrospinal fluid of patients with tuberculosis[J].Pneumonol Alergol Pol,2004,72(3-4):105-110.
[12] Kashyap RS,Kainithla RP,Satpute RM,et al.Demonstration of IgG antibodies to 30 Kd protein antigen in CSF for diagnosis of tuberculous meningitis by antibody capturingELISA[J].Neurol India,2004,52 (3) :359-362.
[13]王仲元,陈红兵,何秀云,等.斑点酶免疫渗滤法抗体检测对结核性脑膜炎诊断价值的评估[J].北京医学,2005,27(2):120-121.
[14] Quan C, Lu CZ, Qiao J,et al.Comparative evaluation of early diagnosis of tuberculous meningitis by different assays[J].J Clin Microbiol, 2006,44(9):3160-3166.
[15] Restrepo BI, Pino PA, Volcy M,et al.Interpretation of mycobacterial antibodies in the cerebrospinal fluid of adults with tuberculous meningitis[J].Trop Med Int Health, 2008 ,13(5):653-658.
[16] Luca MC, Petrovici CM, Va?? A,et al.Gamma interferon testing in blood and cerebrospinal fluid--rapid method for the diagnosis of tuberculous meningitis[J].Rev Med Chir Soc Med Nat Iasi, 2008 ,112(1):108-110.
[17] Thomas MM; Hinks TS; Raghuraman S,et al. Rapid diagnosis of Mycobacterium tuberculosis meningitis by enumeration of cerebrospinal fluid antigen-specific T-cells[J]. Int J Tuberc Lung Dis, 2008 ,12(6): 651-657.
[18] Murakami S, Takeno M, Oka H,et al.Diagnosis of Tuberculous Meningitis Due to Detection of ESAT-6-Specific Gamma Interferon Production in Cerebrospinal Fluid Enzyme-Linked Immunospot Assay[J].Clinical and Vaccine Immunol,2008,15(5) 897-899.
[19] Bordin P,Rosenkrands I,Andersen P,et al.ESAT-6 proteins: protective antigens and virulence factors [J]. Trends Microbiol,2004,12(11):500-508.
[20] Losi, M, Bossink A, Codecasa L, et al. Use of a T-cell interferon-gamma release assay for the diagnosis of tuberculous pleurisy[J]. Eur Respir J, 2007, 30(6):1173-117
[2] Thwaites GE, Nguyen DB,Nquyen HD,et al. Dexamethasone for the treatment of tuberculous meningitis in adolescents and adults. N Engl J Med, 2004, 351(17): 1741-1751.
[3] Vinnard C, Macgregor RR.Tuberculous meningitis in HIV-infected individuals. Curr HIV/AIDS Rep, 2009 ,6(3):139-45.
[4] American Thoracic Society and Centers for Disease Control and Prevention.2000. Targeted tuberculin testing and treatment of latent tuberculosis infection. Am. J. Respir. Crit. Care Med. 161:S221–S247.
[5] Kulkarni SP, Jaleel MA, Kadival GV. Evaluation of an in house developed PCR for the diagnosis of tuberculous meningitis in Indian children. J Med Microbiol, 2005, 54(Pt4): 369-373.
[6] Haldar S, Sharma N, Gupta VK, et al. Efficient diagnosis of tuberculous meningitis by detection of Mycobacterium tuberculosis DNA in cerebrospinal fluid filtrates using PCR.J Med Microbiol, 2009,58(Pt 5):616-624.
[7] Kashyap RS,Kainthla RP,Biswas SK,et al.Rapid diagnosis of tuberculous meningitis using the simple Dot ELISA method.Med Sci Monit, 2003, 9(11): MT123-126.
[8] Kashyap RS, Dobos KM, Belisle JT,et al.Demonstration of components of antigen 85 complex in cerebrospinal fluid of tuberculous meningitis patients.Clin Diagn Lab Immunol ,2005,12 (6) :752-758.
[9] Quan C, Lu CZ, Qiao J,et al.Comparative evaluation of early diagnosis of tuberculous meningitis by different assays.J Clin Microbiol, 2006, 44(9): 3160-3166.
[10] Losi, M, Bossink A, Codecasa L, et al. Use of a T-cell interferon-gamma release assay for the diagnosis of tuberculous pleurisy. Eur Respir J, 2007, 30(6): 1173-1179.
[11] Mathai A,Radhakrishnan VV,Sarada C,et al.Detection of heat stable mycobacterial antigen in cerebrospinal fluid by Dot-Immunobinding assay. Neurol India, 2003 , 51(1):52-54.
[12] Demkow U,Filewska M,Bialas B,et al.Antimycobacterial antibody level in pleural pericardial and cerebrospinal fluid of patients with tuberculosis.Pneumonol Alergol Pol, 2004, 72(3-4):105-110.
[13] Kashyap RS,Kainithla RP,Satpute RM,et al. Demonstration of IgG antibodies to 30 Kd protein antigen in CSF for diagnosis of tuberculous meningitis by antibody capturing ELISA. Neurol India, 2004,52 (3) :359-362.
[14] Quan C, Lu CZ, Qiao J,et al. Comparative evaluation of early diagnosis of tuberculous meningitis by different assays. J Clin Microbiol, 2006, 44(9): 3160-3166.
[15] Luca MC, Petrovici CM, Va?? A, et al.Gamma interferon testing in blood and cerebrospinalfluid--rapid method for the diagnosis of tuberculous meningitis. Rev Med Chir Soc Med Nat Iasi, 2008 ,112(1):108-110.
[16] Thomas MM, Hinks TS, Raghuraman S, et al. Rapid diagnosis of Mycobacterium tuberculosis meningitis by enumeration of cerebrospinal fluid antigen-specific T-cells. Int J Tuberc Lung Dis, 2008 ,12(6): 651-657.
[17] Murakami S, Takeno M, Oka H,et al. Diagnosis of Tuberculous Meningitis Due to Detection of ESAT-6-Specific Gamma Interferon Production in Cerebrospinal Fluid Enzyme-Linked Immunospot Assay.Clinical and Vaccine Immunol,2008,15(5) 897-899.
[18] Srivastava KL, Bansal M, Gupta S,et al.Diagnosis of tuberculous meningitis by detection of antigen and antibodies in CSF and sera. Indian Pediatr. 1998, 35(9): 841-50.
[19] Restrepo BI, Pino PA, Volcy M, et al.Interpretation of mycobacterial antibodies in the cerebrospinal fluid of adults with tuberculous meningitis.Trop Med Int Health, 2008, 13(5):653-658.
[20] Venkatesh K, Parija SC, Mahadevan S,et al.Reverse passive haemagglutination (RPHA) test for detection of mycobacterial antigen in the cerebrospinal fluid for diagnosis of tubercular meningitis. Indian J Tuberc,2007,54(1):41-48.
[21] Sumi MG, Mathai A, Reuben S, et al.Immunocytochemical method for early laboratory diagnosis of tuberculous meningitis. Clin Diagn Lab Immunol, 2002, 9(2):344-347.
[22] Renshaw PS,Panagiotidou P,Whelan A,et al.Conclusive evidence that the major T-cell antigens of the Mycobacterium tuberculosis complex ESAT-6 and CFP-10 form a tight,1:1 complex and characterization of the structural properties of ESAT-6, CFP-10,and the ESAT-6*CFP-10 complex.Implications for pathogenesis and virulence. J Biol Chem, 2002, 277:21598–21603.
[23] Converse, S E, and JS. A protein secretion pathway critical for Mycobacterium tuberculosis virulence is conserved and functional in Mycobacterium smegmatis. J Bacteriol, 2005, 187:1238–1245.
[24] Lewis KN, Liao R, Guinn K M,et al. Deletion of RD1 from Mycobacterium tuberculosis mimics bacille Calmette-Guerin attenuation. J Infect Dis, 2003, 187:117–123.
[25] Amoudy HA, Al-Turab MB, Mustafa AS. Identification of transcriptionally active open reading frames within the RD1 genomic segment of Mycobacterium tuberculosis. Med Princ Pract, 2006, 15:137–144.
[26] van Pinxteren LA, Ravn P, Agger EM, et al. Diagnosis of tuberculosis based on the two specific antigens ESAT-6 and CFP10. Clin Diagn Lab Immunol, 2000, 7(2): 155-160.
[27] Ahuja GK, Mohan KK, Prasad K, Behari M.Diagnostic criteria for tuberculous meningitis and their validation. Tuber Lung Dis, 1994, 75(2):149-152.
[28] Seth R, Sharma U.Diagnostic criteria for Tuberculous Meningitis. Indian J Pediatr, 2002, 69(4):299-303.
[29] Miceli, I., I. N. de Kantor, et al.Evaluation of the effectiveness of BCG vaccination using the case-control method in Buenos Aires, Argentina. Int. J. Epidemiol. 1988, 17:629-634.
[30] Cicero R, Olivera H, Hernández-Solis A,et al.Frequency of Mycobacterium bovis as an etiologic agent in extrapulmonary tuberculosis in HIV-positive and -negative Mexican patients. Eur J Clin Microbiol Infect Dis, 2009, 28(5):455-460.
[31] Shah, N. P., A. Singhal, et al. Occurrence of overlooked zoonotic tuberculosis: detection of Mycobacterium bovis in human cerebrospinal fluid. J. Clin. Microbiol, 2006, 44:1352-1358.
[32] Ritacco V, de Kantor IN.Simultaneous detection of Mycobacterium bovis and Mycobacterium tuberculosis in human cerebrospinal fluid. J Clin Microbiol. 2007, 45(2): 684.
[33] Thwaites G,Caws M,Chau TT,et al.Relationship between Mycobacterium tuberculosis genotype and the clinical phenotype of pulmonary and meningeal tuberculosis. J Clin Microbiol, 2008, 46(4):1363-1368.
[34]王维治,郭玉璞,宿英英.神经病学.人民卫生出版社,2006,1 :670-675.
[35] Flesch I, Gefferth C.X-ray examination of the ventricular system of children suffering from tuberculous meningitis. Paediatr Danub, 1949, 5(3-4):158-168.
[36] Yarami? A, Bükte Y, Katar S, et al.Chest computerized tomography scan findings in 74 children with tuberculous meningitis in southeastern Turkey. Turk J Pediatr, 2007, 49(4): 365-369.
[37] Andronikou S, Wieselthaler N, Smith B,et al.Value of early follow-up CT in paediatric tuberculous meningitis. Pediatr Radiol, 2005, 35(11):1092-1099.
[38] van der Merwe DJ, Andronikou S, Van Toorn R, et al. Brainstem ischemic lesions on MRI in children with tuberculous meningitis: with diffusion weighted confirmation. Childs Nerv Syst, 2009, 25(8):949-954.
[39] Pienaar M, Andronikou S, van Toorn R.MRI to demonstrate diagnostic features and complications of TBM not seen with CT. Childs Nerv Syst, 2009, 25(8):941-947.
[40] Chitre PS, Tullu MS, Sawant HV, et al. Co-occurrence of intracerebral tuberculoma with lumbar intrame dullary tuberculoma. J Child Neurol, 2009, 24(5):606-609.
[41] Xiong CH, Liang XF, Wang HQ. A systematic review on the protective efficacy of BCG against children tuberculosis meningitis and millet tuberculosis. Zhongguo Ji Hua Mian Yi, 2009, 15(4):358-362.
[42] Wasay M, Ajmal S, Taqui AM, et al. Impact of Bacille Calmette-Guerin Vaccination on Neuroradiological Manifestations of Pediatric Tuberculous Meningitis. J Child Neurol, 2009, 30(5):60-63.
[43] Dural AT, Johnson PC, Wagner A. The utility of direct acid-fast stain of cerebrospinal fluid for the rapid diagnosis of tuberculous meningitis in patients with and without AIDS. J Gen Intern Med, 1997,12(4):259.
[44] Radhakrishnan VV, Mathai A, Thomas M. Correlation between culture of Mycobacterium tuberculosis and antimycobacterial antibody in lumbar, ventricular and cisternal cerebrospinal fluids of patients with tuberculous meningitis. Indian J Exp Biol, 1991, 29(9): 845-848.
[45] Krishnan VV,Mathai A.Isolation of two antigens from the culture filtrates of Mycobacterium tuberculosis and their applications in the laboratory diagnosis of the tuberculous meningitis.Med Microbiol Immunol,1991,180(2):101-107.
[46] Baveja CP, Gumma V, Jain M, et al.Newer methods over the conventional diagnostic tests for tuberculous meningitis: do they really help? Trop Doct, 2009, 39(1): 18-20.
[47]赵钢.脑脊液细胞学研究的现状和展望.神经损伤与功能重建,2006,1(3):168-170.
[48] Katti MK.Immunodiagnosis of tuberculous meningitis: rapid detection of mycobacterial antigens in cerebrospinal fluid by reverse passive hemagglutination assay and their characterization by Western blotting. FEMS Immunol Med Microliol, 2001, 31(1):59-64 .
[49] Tiwari RP, Garg SK, Bharmal RN,et al.Rapid liposomal agglutination card test for the detection of antigens in patients with active tuberculosis.Int J Tuberc Lung Dis, 2007, 11(10): 1143-1151.
[50] Samuel AM,Kadival GV,Irani S,et al.A sensitive and specific method for diagnosis of tubercular meningitis. Indian J Med Res, 1983, 77:752-757.
[51] Sada E, Ruiz-Palacios G M, López-Vidal Y, et al. Detection of mycobacterial antigens in cerebrospinal ?uid of patients with tuberculous meningitis by enzyme-linked immunosorbent assay.Lancet.1983,2: 651–652.
[52] Krambovitis E, McIllmurray M B, Lock P E, et al. Rapid diagnosis of tuberculous meningitis by latex particle agglutination.Lancet.1984,2: 1229–1231.
[53] Kadival G V, Mazarelo T B, Haparas S D. Sensitivity and specificity of enzyme linked immunosorbent assay in the detection of antigen in tuberculous meningitis cerebrospinal ?uids. J Clin Microbiol. 1986,23: 901–904.
[54] Murakami S, Takeno M, Oka H, et al.Diagnosis of tuberculous meningitis due to detection of ESAT-6-specific gamma interferon production in cerebrospinal fluid enzyme-linked immunospot assay.Clin Vaccine Immunol,2008,15(5):897-899.
[55] Losi, M, Bossink A, Codecasa L, et al. Use of a T-cell interferon-gamma release assay for the diagnosis of tuberculous pleurisy. Eur Respir J, 2007, 30(6): 1173-1179.
[56] Cole ST, Brosch R, Parkhill J, et al. Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence.Nature,1998,393:537–544.
[57] Hart PD, Sutherland I. BCG and vole bacillus vaccines in the prevention of tuberculosis in adolescence andearly adult life. Br Med J, 1977, 2:293–295.
[58] Sorensen AL, Nagai S, Houen G,et al.Purification and characterization of a low-molecular-mass T-cell antigen secreted by Mycobacterium tuberculosis. Infect Immun, 1995,63:1710–1717.
[59] Berthet FX, Rasmussen PB, Rosenkrands I, et al. Mycobacterium tuberculosis operon en-coding ESAT-6 and a novel low-molecular-mass culture filtrate protein (CFP-10). Microbiology, 1998,144:3195–3203.
[60] Brodin P, Rosenkrands I, Andersen P, et al.ESAT-6 proteins: protective antigens and virulence factors? Trends Microbiol, 2004,12:500–508.
[61] Pym AS, Brodin P, Majlessi L,et al.Recombinant BCG exporting ESAT-6 confers enhanced protection against tuberculosis. Nat Med, 2003,9:533–539.
[62] Guinn KM, Hickey MJ, Mathur SK, et al. Individual RD1-region genes are required for export of ESAT-6/CFP-10 and for virulence of Mycobacterium tuberculosis. Mol Microbiol, 2004, 51:359–370.
[63] Gao LY, Guo S, McLaughlin B, et al.A mycobacterial virulence gene cluster extending RD1 is required for cytolysis, bacterial spreading and ESAT-6 secretion. Mol Microbiol, 2004, 53:1677–1693.
[64] Pym AS, Brodin P, Majlessi L, et al. Recombinant BCG exporting ESAT-6 confers enhanced protection against tuberculosis. Nat Med, 2003, 9:533–539.
[65] Lightbody KL, Renshaw PS, Collins ML,et al.Characterisation of complex formation between members of the Mycobacterium tuberculosis complex CFP-10/ESAT-6 protein family:towards an understanding of the rules governing complex formation and thereby functional flexibility. FEMS Microbiol Lett, 2004,238:255–262.
[66] Renshaw PS, Panagiotidou P, Whelan A,et al.Conclusive evidence that the major T-cell antigens of the Mycobacterium tuberculosis complex ESAT-6 and CFP-10 form a tight, 1:1 complex and characterization of the structural properties of ESAT-6, CFP-10,and the ESAT-6*CFP-10 complex. Implications for pathogenesis and virulence. J Biol Chem, 2002,277:21598–21603.
[67] Papps M G, Ajkowski R, Hockmeyer W T. Dot-enzyme-linked immunosorbent assay (dot-ELISA): a micro technique for rapid diagnosis of visceral leishmaniasis. J Immunol Methods, 1983, 64(1-2): 205-214.
[68] Colangeli R, Spencer JS, Bifani P,et al. MTSA-10, the product of the Rv3874 gene of Mycobacterium tuberculosis, elicits tuberculosis-specific, delayed-type hypersensitivity in guinea pigs. Infect Immun, 2000, 68:990–993.
[69] Sahn SA. Pleural thickening, trapped lung, and chronic empyemaas sequelae of tuberculous pleural effusion: don’t sweat the pleuralthickening. Int J Tuberc Lung Dis, 2002, 6: 461-464.
[70] Shenai SC ,Rodrigues, A P Mehta,et al. Evaluation of a new phage Amplification technology for rapid diagnosis of tuberculosis. Indian J. Med. Microbiol, 2002, 20:194–199.
[71] Harboe M, Oettinger T, Wiker HG, et al. Evidence for occurrence of the ESAT-6 protein in Mycobacterium tuberculosis and virulent Mycobacterium bovis and for its absence in Mycobacterium bovis BCG. Infect Immun, 1996, 64:16–22.
[72]全国结核病流行病学抽样调查技术指导组.第四次全国结核病流行病学抽样调查报告.中华结核和呼吸杂志,2002,25(1):6-10.
[1] Thwaites GE, Chau TT,Stepniewska K,et al. Diagnosis of adult tuberculous meningitis by use of clinical and laboratory features[J]. Lancet, 2002,360(9342):1287-1292.
[2] Thwaites GE, Nguyen DB,Nquyen HD,et al. Dexamethasone for the treatment of tuberculous meningitis in adolescents and adults[J]. N Engl J Med, 2004,351(17):1741-1751.
[3] Kulkarni SP, Jaleel MA, Kadival GV. Evaluation of an in-house-developed PCR for the diagnosis of tuberculous meningitis in Indian children[J]. J Med Microbiol, 2005, 54(Pt4):369-373.
[4] Sumi MG, Mathai A, Reuben S, et al.Immunocytochemical method for early laboratory diagnosis of tuberculous meningitis[J]. Clin Diagn Lab Immunol, 2002, 9(2):344-347.
[5] Kashyap RS,Kainthla RP,Biswas SK,et al.Rapid diagnosis of tuberculous meningitis using the simple Dot ELISA method[J].Med Sci Monit,2003,9(11):MT123-126.
[6] Mathai A,Radhakrishnan VV,Sarada C,et al.Detection of heat stable mycobacterial antigen in cerebrospinal fluid by Dot-Immunobinding assay [J].Neurol India,2003 ,51(1):52-54.
[7] Kashyap RS, Dobos KM, Belisle JT,et al.Demonstration of components of antigen 85 complex in cerebrospinal fluid of tuberculous meningitis patients[J].Clin Diagn Lab Immunol ,2005,12 (6) :752-758.
[8] Venkatesh K, Parija SC, Mahadevan S,et al.Reverse passive haemagglutination (RPHA) test for detection of mycobacterial antigen in the cerebrospinal fluid for diagnosis of tubercular meningitis [J].Indian J Tuberc,2007,54(1):41-48.
[9]丁晓旭,薛承岩.脑脊液和血液标本检测阿拉伯糖甘露糖脂G抗体诊断结核性脑膜炎的价值[J ].临床荟萃,2007,22 (4) :284-286.
[10]李永刚,杨昆胜,晋华文,等.脂阿拉伯甘露聚糖诊断结核性脑膜炎的评价[J].中国防痨杂志,2005, 27(5):316-319 .
[11] Demkow U,Filewska M,Bialas B,et al.Antimycobacterial antibody level in pleural , pericardial and cerebrospinal fluid of patients with tuberculosis[J].Pneumonol Alergol Pol,2004,72(3-4):105-110.
[12] Kashyap RS,Kainithla RP,Satpute RM,et al.Demonstration of IgG antibodies to 30 Kd protein antigen in CSF for diagnosis of tuberculous meningitis by antibody capturingELISA[J].Neurol India,2004,52 (3) :359-362.
[13]王仲元,陈红兵,何秀云,等.斑点酶免疫渗滤法抗体检测对结核性脑膜炎诊断价值的评估[J].北京医学,2005,27(2):120-121.
[14] Quan C, Lu CZ, Qiao J,et al.Comparative evaluation of early diagnosis of tuberculous meningitis by different assays[J].J Clin Microbiol, 2006,44(9):3160-3166.
[15] Restrepo BI, Pino PA, Volcy M,et al.Interpretation of mycobacterial antibodies in the cerebrospinal fluid of adults with tuberculous meningitis[J].Trop Med Int Health, 2008 ,13(5):653-658.
[16] Luca MC, Petrovici CM, Va?? A,et al.Gamma interferon testing in blood and cerebrospinal fluid--rapid method for the diagnosis of tuberculous meningitis[J].Rev Med Chir Soc Med Nat Iasi, 2008 ,112(1):108-110.
[17] Thomas MM; Hinks TS; Raghuraman S,et al. Rapid diagnosis of Mycobacterium tuberculosis meningitis by enumeration of cerebrospinal fluid antigen-specific T-cells[J]. Int J Tuberc Lung Dis, 2008 ,12(6): 651-657.
[18] Murakami S, Takeno M, Oka H,et al.Diagnosis of Tuberculous Meningitis Due to Detection of ESAT-6-Specific Gamma Interferon Production in Cerebrospinal Fluid Enzyme-Linked Immunospot Assay[J].Clinical and Vaccine Immunol,2008,15(5) 897-899.
[19] Bordin P,Rosenkrands I,Andersen P,et al.ESAT-6 proteins: protective antigens and virulence factors [J]. Trends Microbiol,2004,12(11):500-508.
[20] Losi, M, Bossink A, Codecasa L, et al. Use of a T-cell interferon-gamma release assay for the diagnosis of tuberculous pleurisy[J]. Eur Respir J, 2007, 30(6):1173-117