Please wait a minute...
浙江大学学报（医学版）
浙江大学学报（医学版）  2016, Vol. 45 Issue (3): 243-248    DOI: 10.3785/j.issn.1008-9292.2016.05.04
艾滋病专题     
艾滋病合并分枝杆菌感染患者分枝杆菌菌种鉴定
宋炜1, 刘莉1, 卢洪洲1,2
1. 上海市公共卫生临床中心感染一科, 上海 201508;
2. 复旦大学附属华山医院感染病科, 上海 200040
Identification of Mycobacteria isolated from AIDS patients
SONG Wei1, LIU Li1, LU Hongzhou1,2
1. Department of Infectious Disease, Shanghai Public Health Clinical Center, Shanghai 201508, China;
2. Department of Infectious Disease, Huashan Hospital, Fudan University, Shanghai 200040, China
全文: PDF(1004 KB)  
摘要: 

目的：对艾滋病合并分枝杆菌感染患者菌株标本进行菌种鉴定，评价MPB64抗原胶体金法快速检测试剂盒鉴别分枝杆菌的价值。方法：对艾滋病患者临床标本中培养出的140株分枝杆菌菌株用16SrDNA测序的方法进行菌种鉴定，分析艾滋病患者中常见非结核性分枝杆菌（NTM）菌种，同时对这些标本用MPB64抗原免疫胶体金法进行菌株鉴定，分析后者的敏感性和特异性。结果：以16SrDNA测序结果为金标准，MPB64抗原胶体金法快速检测区分结核分枝杆菌（MTB）与NTM的敏感性和特异性分别为96.00%和96.84%。共有114份标本（114/140，81.43%）获得明确鉴定结果，其中50份（43.86%）为MTB，63份标本（55.26%）鉴定为NTM。最常见的NTM菌种为戈登分枝杆菌、鸟分枝杆菌和堪萨斯分枝杆菌。结论：MPB64抗原胶体金法检测分枝杆菌菌种具有较好的特异性，且快速、经济、简便，适合用于临床对MTB及NTM的初步鉴别。艾滋病合并结核患者中NTM的比例较高，需进行菌种鉴定以指导临床治疗。
关键词 ： 获得性免疫缺陷综合征,  分枝杆菌,结核/分离和提纯,  分枝杆菌属/分类,  寡核苷酸序列分析,  细菌蛋白质类/遗传学,  胶体金,  试剂盒,诊断    
Abstract：

Obstract: To identify Mycobacteria isolated from AIDS patients and to evaluate the diagnostic value of MBP64 antigen Colloidal Gold Diagnostic Kit in distinguishing Mycobacterium tuberculosis(MTB) and Nontuberculous mycobacteria (NTM). Methods: A total of 140 Mycobacteria isolates from AIDS patients were collected and identified by using 16SrDNA PCR assay as well as immune colloidal gold technique (MBP64 antigen Colloidal Gold Diagnostic Kit). The sensitivity and specificity of Colloidal Gold method was analyzed. Results: With the results of 16SrDNA PCR assay as gold standard, the sensitivity and specificity of Colloidal Gold method in distinguishing MTB and NTM were 96.00% and 96.84%, respectively. Among 140 samples, 114 were identified by 16SrDNA PCR assay, among which 50 (43.86%) were identified as MTB, and 63 (55.26%) were identified as NTM. The most popular NTM in AIDS patients were Mycobacterium gordonae, Mycobacterium avium and Mycobacterium kansasii. Conclusion: NTM is of high prevalence in AIDS patients, and MBP64 antigen Colloidal Gold Diagnostic Kit can be used as an effective technique in distinguishing NTM and MTB.
Key words： Acquired immunodeficiency syndrome    Mycobacterium tuberculosis/isolation &    purification    Mycobacterium/classification    Oligonucleotide array sequence analysis    Bacterial proteins/genetics    Gold colloid    Reagent kits, diagnostic
收稿日期: 2016-01-10
CLC:  R446.5  
基金资助:

国家十二五科技重大专项（2012ZX10001003，2012ZX09303013）；国家高技术研究发展计划（863计划）（2014AA021403）
通讯作者: 卢洪洲(1966-),男,博士,教授,主任医师,博士生导师,主要从事艾滋病基础与临床研究;E-mail:luhongzhou@fudan.edu.cn;http://orcid.org/0000-0002-8308-5534     E-mail: luhongzhou@fudan.edu.cn
作者简介: 宋炜(1982-),男,学士,主治医师,从事艾滋病及相关机会性感染的研究;E-mail:songwei@shaphc.org;http://orcid.org/0000-0002-0983-0823
服务　　
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章　　

引用本文:

宋炜 等. 艾滋病合并分枝杆菌感染患者分枝杆菌菌种鉴定[J]. 浙江大学学报（医学版）, 2016, 45(3): 243-248.
SONG Wei, LIU Li, LU Hongzhou. Identification of Mycobacteria isolated from AIDS patients. Journal of ZheJiang University(Medical Science), 2016, 45(3): 243-248.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.1008-9292.2016.05.04      或      http://www.zjujournals.com/xueshu/med/CN/Y2016/V45/I3/243

[1] World Health Organization. WHO global tuberculosis report 2015[M]. Geneva: World Health Organization,2015:13.
[2] World Health Organization. Multidrug and extensively drug-resistant TB (M/XDR-TB): 2010 global report on surveillance and response[M]. Geneva:World Health Organization,2012:17-19.
[3] JOHNSON, J L, ELLNER, J J. Tropical infectious diseases: principles, pathogens and practice[M]. Philadelphia:Churchill Livingstone,2011:228-247.
[4] WOLINSKY E. Mycobacterial diseases other than tuberculosis[J]. Clin Infect Dis, 1992,15(1):1-10.
[5] HERZMANN C, LANGE C. Infections with Non-tuberculous mycobacteria and HIV[J]. Dtsch Med Wochenschr,2010,135(23):1192-1197.
[6] KAWANA M, STARR R S, TASHIMA K T, et al. Spontaneous perforation of the terminal ileum in an AIDS patient on highly active antiretroviral therapy with disseminated non-tuberculous mycobacterial infection[J]. Int J Infect Dis,2008,12(6):603-606.
[7] MIGUEZ-BURBANO M J, FLORES M, ASHKIN D, et al. Non-tuberculous mycobacteria disease as a cause of hospitalization in HIV-infected subjects[J]. Int J Infect Dis, 2006,10(1):47-55.
[8] MIGUEZ-BURBANO M J, SHOR-POSNER G, HADRIGAN S. Non-tuberculous mycobacteria in HIV-infected patients: geographic, behavioural, and immunological factors[J]. Lancet Infect Dis,2005,5(7):394-395.
[9] EL-SOLH A A, NOPPER J, ABDUL-KHOUDOUD M R, et al. Clinical and radiographic manifestations of uncommon pulmonary nontuberculous mycobacterial disease in AIDS patients[J]. Chest,1998,114(1):138-145.
[10] POKAM B T, ASUQUO A E. (2012) Acid-fast bacilli other than Mycobacteria in tuberculosis patients receiving directly observed therapy short course in cross river state, Nigeria[J]. Tuberc Res Treat,2012,2012:301056.
[11] AL JARAD N, DEMERTZIS P, JONES D J, et al. Comparison of characteristics of patients and treatment outcome for pulmonary non-tuberculous mycobacterial infection and pulmonary tuberculosis[J]. Thorax,1996,51(2):137-139.
[12] AFGHANI B, LIEBERMAN J M, DUKE M B, et al. Comparison of quantitative polymerase chain reaction, acid fast bacilli smear, and culture results in patients receiving therapy for pulmonary tuberculosis[J]. Diagn Microbiol Infect Dis,1997,29(2):73-79.
[13] CRUCIANI M, SCARPARO C, MALENA M, et al. Meta-analysis of BACTEC MGIT 960 and BACTEC 460 TB, with or without solid media, for detection of Mycobacteria[J]. J Clin Microbiol,2004,42(5):2321-2325.
[14] KATILA M L, KATILA P, ERKINJUNTTI-PEKKANEN R. Accelerated detection and identification of Mycobacteria with MGIT 960 and COBAS AMPLICOR systems[J]. J Clin Microbiol,2000,38(3):960-964.
[15] CLOUD J L, NEAL H, ROSENBERRY R, et al. Identification of Mycobacterium spp. by using a commercial 16S ribosomal DNA sequencing kit and additional sequencing libraries[J]. J Clin Microbiol,2002,40(2):400-406.
[16] REDDINGTON K, O'GRADY J, DORAI-RAJ S, et al. A novel multiplex real-time PCR for the identification of Mycobacteria associated with zoonotic tuberculosis[J/OL]. PLoS One,2011,6(8):e23481.
[17] MOURE R, MARTIN R, ALCAIDE F. Effectiveness of an integrated real-time PCR method for detection of the Mycobacterium tuberculosis complex in smear-negative extrapulmonary samples in an area of low tuberculosis prevalence[J]. J Clin Microbiol,2012,50(2):513-515.
[18] LING D I, FLORES L L, RILEY L W, et al. Commercial nucleic-acid amplification tests for diagnosis of pulmonary tuberculosis in respiratory specimens: meta-analysis and meta-regression[J/OL]. PLoS One,2008,3(2): e1536.
[19] WANG L, CHENG S, XU M, et al. Model collaboration between hospitals and public health system to improve tuberculosis control in China[J]. Int J Tuberc Lung Dis,2009,13(12): 1486-1492.
[20] VAN SOOLINGEN D, HWEMANS P W, DE HAAS P E, et al. Occurrence and stability of insertion sequences in Mycobacterium tuberculosis complex strains: evaluation of an insertion sequence-dependent DNA polymorphism as a tool in the epidemiology of tuberculosis[J]. J Clin Microbiol,1991,29(11):2578-2586.
[21] LAN R, YANG C, LAN L, et al. Mycobacterium tuberculosis and Non-tuberculous mycobacteria isolates from HIV-infected patients in Guangxi, China[J]. Int J Tuberc Lung Dis,2011,15(12):1669-1675.
[22] KHATTER S, SINGH U B, ARORA J, et al. Mycobacterial infections in human immuno-deficiency virus seropositive patients: role of Non-tuberculous mycobacteria[J]. Indian J Tuberc,2008,55(1): 28-33.
[23] BUIJTELS P C, VAN-DER-SANDE M A, DE-GRAAFF C S, et al. Non-tuberculous mycobacteria, Zambia[J]. Emerg Infect Dis, 2009, 15(2):242-249.
[24] OLSON E S, SIMPSON A J, NORTON A J, et al. Not everything acid fast is Mycobacterium tuberculosis: a case report[J]. J Clin Pathol,1998,51(7):535-536.
[25] NAGAI S, WIKER H G, HARBOE M, et al. Isolation and partial characterization of major protein antigens in the culture fluid of Mycobacterium tuberculosis[J]. Infect Immun,1991,59(1):372-382.
[26] ABE C, HIRANO K, TOMIYAMA T. Simple and rapid identification of Mycobacterium tuberculosis complex by immunochromatographic assay using anti-MPB64 monoclonal antibodies[J]. J Clin Microbiol,1999,37(11):3693-3697.
[27] HILLEMANN D, RVSCH-GERDES S, RICHTER E. Application of the Capilia TB assay for culture confirmation of Mycobacterium tuberculosis complex isolates[J]. Int J Tuberc Lung Dis,2005,9(12):1409-1411.
[28] MUYOYETA M, DE HAAS P E, MUELLER D H, et al. Evaluation of the Capilia TB assay for culture confirmation of Mycobacterium tuberculosis infections in Zambia and South Africa[J]. J Clin Microbiol,2010,48(10):3773-3775.
[29] YAMAGUCHI R, MATSUO K, YAMAZAKI A, et al. Cloning and characterization of the gene for immunogenic protein MPB64 of Mycobacterium bovis BCG[J]. Infect Immun,1989,57(1):283-288.
[30] TAMADA Y, KANDA S, YOSHIDOME A, et al. Diagnosis of active tuberculosis using MPB64, a specific antigen of Mycobacterium bovis[J]. Microbiol Immunol,2012,56(11):740-747.
[31] ZAKOWSKI P, FLIGIEL S, BERLIN G W, et al. Disseminated Mycobacterium avium-intracellulare infection in homosexual men dying of acquired immunodeficiency[J]. JAMA,1982,248(22):2980-2982.
[32] HOSBURGH C R, HAVLIK J A, ELLIS D A, et al. Survival of patients with acquired immune deficiency syndrome and disseminated Mycobacterium avium complex infection with and without antimycobacterial chemotherapy[J]. Am Rev Respir Dis,1991,144(3 Pt 1):557-559.
[33] NIGHTINGALE S D, BYRD L T, SOUTHERN P M, et al. Incidence of Mycobacterium avium-intracellulare complex bacteremia in human immunodeficiency virus-positive patients[J]. J Infect Dis,1992,165(6):1082-1085.
[34] TORTOLI E. Impact of genotypic studies on mycobacterial taxonomy: the new mycobacteria of the 1990s[J]. Clin Microbiol Rev,2003,16(2):319-354.
[1] 沈银忠 等. 人类免疫缺陷病毒暴露前预防的应用与挑战[J]. 浙江大学学报（医学版）, 2016, 45(3): 221-227.
[2] 王炬峰 等. 高效抗逆转录病毒治疗对人类免疫缺陷病毒感染者骨代谢的影响[J]. 浙江大学学报（医学版）, 2016, 45(3): 228-235.
[3] 林海江 等. 人类免疫缺陷病毒共感染夫妻艾滋病相关危险行为网络及分子流行病学研究[J]. 浙江大学学报（医学版）, 2016, 45(3): 236-242.
[4] 孙建军等. 《艾滋病诊疗指南第三版(2015版)》更新解读[J]. 浙江大学学报（医学版）, 2015, 44(6): 597-602.
[5] 吴丽娟，靳昌忠，白石，梁勇，吴南屏. 合并病毒性肝炎HIV感染患儿接受高效抗逆转录酶病毒治疗后肝功能变化[J]. 浙江大学学报（医学版）, 2014, 43(2): 180-186.
[6] . 新疆女性性工作者艾滋病暴露前预防用药使用意愿调查[J]. 浙江大学学报（医学版）, 2011, 40(3): 281-285.
[7] 孙健,吴南屏,冯婷婷,冯磊,Norbert Brockmeyer. 长期高效抗逆转录病毒治疗对HIV-1感染者体内异常免疫激活和免疫重建的影响[J]. 浙江大学学报（医学版）, 2010, 39(6): 618-622.
[8] 杨冰，黄乐群. 双功能化胶体金纳米探针的制备及其特异性考察[J]. 浙江大学学报（医学版）, 2010, 39(3): 296-304.
[9] 姚航平，靳昌忠，张福杰，冯磊，魏红山，吴灵娇，郜桂菊，Armin Bader，Norbert Brockmeyer，吴南屏. 高效抗逆转录病毒治疗对AIDS患者MCP-1和MSP水平的影响[J]. 浙江大学学报（医学版）, 2007, 36(2): 174-178.
[10] 王媛，罗依惠，严杰. 幽门螺杆菌cagA基因片段原核表达载体的构建及其cagA基因和CagA蛋白及感染者血清抗体的检测[J]. 浙江大学学报（医学版）, 2005, 34(3): 223-229.
[11] 朱彪，吴南屏，Hoxtermann Stefan，Armin Bader，Norbert Brockmeyer. 艾滋病相关的卡波西肉瘤免疫激活状态研究[J]. 浙江大学学报（医学版）, 2003, 32(2): 101-103.
[12] 沈建根;朱永良;董玉娥;程东庆;鲍建芳;邵传森. 斑点免疫金渗滤试验检测淋球菌抗原的应用研究[J]. 浙江大学学报（医学版）, 2000, 29(2): 61-62.
[13] 瞿国伟, 梁毅, 汤德骥, 庞兴光, 郎亚民, 俞运彪. 四例血友病患者感染艾滋病毒后的社会和心理及防治问题[J]. 浙江大学学报（医学版）, 1994, 23(1): 37-38,40.