Please wait a minute...
浙江大学学报（医学版）
Journal of ZheJiang University(Medical Science)  2016, Vol. 45 Issue (3): 243-248    DOI: 10.3785/j.issn.1008-9292.2016.05.04
    
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
Download:   PDF(1004KB)
Export: BibTeX | EndNote (RIS)      

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     
Received: 10 January 2016     
CLC:  R446.5  
  R512.91  
  R517.9  
Service
E-mail this article
Add to my bookshelf
Add to citation manager
E-mail Alert
RSS
Articles by authors
Cite this article:

SONG Wei, LIU Li, LU Hongzhou. Identification of Mycobacteria isolated from AIDS patients. Journal of ZheJiang University(Medical Science), 2016, 45(3): 243-248.

URL:

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


艾滋病合并分枝杆菌感染患者分枝杆菌菌种鉴定

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 
[[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] SHEN Yinzhong, LU Hongzhou. Pre-exposure prophylaxis for HIV: clinical practice and challenge[J]. Journal of ZheJiang University(Medical Science), 2016, 45(3): 221-227.
[2] WANG Jufeng, XU Lijun, HUANG Ying, XIE Yirui, SUN Jia, SU Junwei, ZHU Biao. Impact of long-term highly active antiretroviral therapy on bone metabolism in AIDS patients[J]. Journal of ZheJiang University(Medical Science), 2016, 45(3): 228-235.
[3] LIN Haijiang, SHEN Weiwei, QIU Danhong, ZHOU Sujuan, WU Qionghai, DING Yingying, LIU Xing, HE Na. HIV related risky behavioral network and molecular epidemiology of HIV seroconcordant couples in Taizhou municipality of Zhejiang Province[J]. Journal of ZheJiang University(Medical Science), 2016, 45(3): 236-242.
[4] SU Min, CHEN Jin, BAI Bing, HUANG Yunxiu, WEI Lan, LIU Minyan, CHEN Tingmei. Establishment and preliminary application of detection of Mycobacterium tuberculosis in sputum based on variable number tandem repeat[J]. Journal of ZheJiang University(Medical Science), 2016, 45(1): 61-67.
[5] SUN Jian-jun, LU Hong-zhou. Highlights of the third edition of Chinese guidelines for AIDS diagnosis and treatment(2015)[J]. Journal of ZheJiang University(Medical Science), 2015, 44(6): 597-602.
[6] LU Xiao-yun，et al. Particle settling characteristics of alcohol precipitation mixture of Paeoniae Radix rubra extract and models establishment of settling velocity [J]. Journal of ZheJiang University(Medical Science), 2014, 43(4): 458-465.
[7] WU Li-juan, JIN Chang-zhong, BAI Shi, LIANG Yong, WU Nan-ping. Highly active antiretroviral therapy on liver function in HIV-positive children with HBV/HCV co-infection[J]. Journal of ZheJiang University(Medical Science), 2014, 43(2): 180-186.
[8] SHEN Qian, YU Lu-shan, ZHOU Hui, ZENG Su. Simultaneous determination of madecassoside，asiaticoside and their aglycones in Centella asiatica (L.) urban extracts by RP-HPLC[J]. Journal of ZheJiang University(Medical Science), 2014, 43(2): 135-140.
[9] ZHU Qian-ying, YU Lu-shan, ZHENG Guo-gang, XIE Sheng-gu, TAO Qiao-feng. Enantiomeric separation and impurity determination of valaciclovir hydrochloride[J]. Journal of ZheJiang University(Medical Science), 2014, 43(2): 164-167.
[10] ZHANG Jie, LI Xiu-Juan. Biological characteristics of phage SM1 for Stenotrophomonas maltophilia and its effect in animal infection model[J]. Journal of ZheJiang University(Medical Science), 2013, 42(3): 331-336.
[11] . Extraction and purification of acidic polysaccharide from Moerella iridescens[J]. Journal of ZheJiang University(Medical Science), 2012, 41(5): 569-575.
[12] . Optimization of genomic DNA extraction with magnetic bead- based semi-automatic system[J]. Journal of ZheJiang University(Medical Science), 2012, 41(3): 320-326.
[13] . Acceptability of pre-exposure prophylaxis among female sex workers in Xinjiang[J]. Journal of ZheJiang University(Medical Science), 2011, 40(3): 281-285.
[14] . Enzymatic activities of recombinant human NAMPT and NAMPT (H247A) proteins[J]. Journal of ZheJiang University(Medical Science), 2011, 40(2): 156-162.
[15] . Detection of N. gonorrhoeae Antigen by Dot-immunogold Filtration Assay[J]. Journal of ZheJiang University(Medical Science), 2000, 29(2): 61-62.