Clonal expansion of Mycobacterium tuberculosis isolates and coexisting drug resistance in patients newly diagnosed with pulmonary tuberculosis in Hanoi, Vietnam

BMC Research Notes

Naoto Keicho³  Luu Thi Lien⁶  Pham Huu Thuong⁵  Shinsaku Sakurada⁴  Nguyen Thi-Le Hang¹  Tomoko Kuwahara⁴  Tran Thi-Bich Thuy²  Hiroki Ando⁴  Nguyen Van Hung² 

[1] NCGM-Bach Mai Hospital Medical Collaboration Center, Hanoi, Vietnam;Department of Microbiology, National Lung Hospital, 463 Hoang Hoa Tham, Hanoi, Vietnam;Research Institute of Tuberculosis JATA, Tokyo, Japan;National Center for Global Health and Medicine, Tokyo, Japan;Hanoi Lung Hospital, Hanoi, Vietnam;Hanoi Department of Health, Hanoi, Vietnam

关键词: Vietnam;    Restriction fragment length polymorphism;    Gene mutation;    Isoniazid;    Primary drug resistance;   

Others  :  1140912
DOI  :  10.1186/1756-0500-6-444

 received in 2013-05-03, accepted in 2013-10-29,  发布年份 2013

【 摘 要 】

Background

Newly diagnosed patients without anti-tuberculosis (TB) treatment histories have not often undergone drug susceptibility testing (DST), but have received the standard treatment regimen without information about their DST profiles in many countries with inadequate resources.

Methods

We collected 346 clinical isolates from previously untreated patients with smear-positive active TB in Hanoi, the capital of Vietnam. Of these, 339 were tested for susceptibility to four first-line anti-TB drugs, including isoniazid (INH), rifampicin (RMP), streptomycin (SM), and ethambutol (EMB), using the proportion method. A pyrazinamidase (PZase) test was used to assess pyrazinamide (PZA) resistance. Results of the culture-based drug susceptibility tests were confirmed by those from reverse hybridization-based line probe assays (LiPAs) that detected mutations associated with RMP, INH, PZA, and fluoroquinolone (FQ) resistance. To investigate a diversity of these strains, IS6110-probed restriction fragment length polymorphisms (RFLPs) were analyzed. Nucleotide sequences for furA-katG and fabG1-inhA operons, transcription units responsible for INH resistance, were also determined.

Results

Of the isolates tested, 127 (37.5%) were resistant to at least one of the four drugs, which included 93 (27.4%) isolates that were resistant to INH. RFLP analysis identified four clusters defined by similarity of the band patterns, which accounted for 46.1% of the tested isolates. Among the clustered isolates, 37.7% were resistant to INH, most of which (85.4%) carried a g944c mutation, which causes an S315T amino acid substitution, in the katG gene.

Conclusions

Our results suggest that drug-resistant strains, particularly those with INH resistance characterized by a single mutation, S315T, are spreading in Hanoi, Vietnam. When RMP resistance is combined with this setting, patients are not easily cured by conventional short-term treatment. We will need to carefully monitor these trends and search for the origins and transmission routes of these strains.

【 授权许可】

   
2013 Hung et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

【 参考文献 】

• [1]Lew W, Pai M, Oxlade O, Martin D, Menzies D: Initial drug resistance and tuberculosis treatment outcomes: systematic review and meta-analysis. Ann Intern Med 2008, 149:123-134.
• [2]Kato-Maeda M, Metcalfe JZ, Flores L: Genotyping of Mycobacterium tuberculosis: application in epidemiologic studies. Future Microbiol 2011, 6:203-216.
• [3]Sougakoff W: Molecular epidemiology of multidrug-resistant strains of Mycobacterium tuberculosis. Clin Microbiol Infect 2011, 17:800-805.
• [4]World Health Organization: Global tuberculosis report 2012. 2012. http://apps.who.int/iris/bitstream/10665/75938/1/9789241564502_eng.pdf webcite, WHO/HTM/TB/2012.6
• [5]World Health Organization: Anti-tuberculosis drug resistance in the world, fourth global report. WHO report; 2008. http://whqlibdoc.who.int/hq/2008/WHO_HTM_TB_2008.394_eng.pdf webcite, WHO/HTM/TB/2008.394
• [6]Le TK, Bach KH, Ho ML, Le NV, Nguyen TN, Chevrier D, Guesdon JL: Molecular fingerprinting of Mycobacterium tuberculosis strains isolated in Vietnam using IS6110 as probe. Tuber Lung Dis 2000, 80:75-83.
• [7]Quy HT, Buu TN, Cobelens FG, Lan NT, Lambregts CS, Borgdorff MW: Drug resistance among smear-positive tuberculosis patients in Ho Chi Minh City, Vietnam. Int J Tuberc Lung Dis 2006, 10:160-166.
• [8]Huong NT, Lan NT, Cobelens FG, Duong BD, Co NV, Bosman MC, Kim SJ, van Soolingen D, Borgdorff MW: Antituberculosis drug resistance in the south of Vietnam: prevalence and trends. J Infect Dis 2006, 194:1226-1232.
• [9]Khuê PM, Phuc TQ, Hung NV, Jarlier V, Robert J: Drug resistance and HIV co-infection among pulmonary tuberculosis patients in Haiphong City, Vietnam. Int J Tuberc Lung Dis 2008, 12:763-768.
• [10]Guidelines for surveillance of drug resistance in tuberculosis. 2nd edition. WHO report; 2003. http://whqlibdoc.who.int/publications/2003/9241546336.pdf webcite, WHO/CDS/TB/2003.320
• [11]Wayne LG: Simple pyrazinamidase and urease tests for routine identification of mycobacteria. Am Rev Respir Dis 1974, 109:147-151.
• [12]Sekiguchi J, Nakamura T, Miyoshi-Akiyama T, Kirikae F, Kobayashi I, Augustynowicz-Kopec E, Zwolska Z, Morita K, Suetake T, Yoshida H, Kato S, Mori T, Kirikae T: Development and evaluation of a line probe assay for rapid identification of pncA mutations in pyrazinamide-resistant mycobacterium tuberculosis strains. J Clin Microbiol 2007, 45:2802-2807.
• [13]van Soolingen D, Hermans PW, de Haas PE, Soll DR, van Embden JD: 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 Clin Microbiol 1991, 29:2578-2586.
• [14]Otsuka Y, Parniewski P, Zwolska Z, Kai M, Fujino T, Kirikae F, Toyota E, Kudo K, Kuratsuji T, Kirikae T: Characterization of a trinucleotide repeat sequence (CGG)5 and potential use in restriction fragment length polymorphism typing of Mycobacterium tuberculosis. J Clin Microbiol 2004, 42:3538-3548.
• [15]Rossau R, Traore H, De Beenhouwer H, Mijs W, Jannes G, De Rijk P, Portaels F, Evaluation of the INNO-LiPA Rif: TB assay, a reverse hybridization assay for the simultaneous detection of Mycobacterium tuberculosis complex and its resistance to rifampin. Antimicrob Agents Chemother 1997, 41:2093-2098.
• [16]Ando H, Kondo Y, Suetake T, Toyota E, Kato S, Mori T, Kirikae T: Identification of katG mutations associated with high-level isoniazid resistance in Mycobacterium tuberculosis. Antimicrob Agents Chemother 2010, 54:1793-1799.
• [17]Ando H, Mitarai S, Kondo Y, Suetake T, Kato S, Mori T, Kirikae T: Evaluation of a line probe assay for the rapid detection of gyrA mutations associated with fluoroquinolone resistance in multidrug-resistant Mycobacterium tuberculosis. J Med Microbiol 2011, 60:184-188.
• [18]van Embden JD, Cave MD, Crawford JT, Dale JW, Eisenach KD, Gicquel B, Hermans P, Martin C, McAdam R, Shinnick TM, Small PM: Strain identification of Mycobacterium tuberculosis by DNA fingerprinting: recommendations for a standardized methodology. J Clin Microbiol 1993, 31:406-409.
• [19]Warren RM, Richardson M, Sampson SL, van der Spuy GD, Bourn W, Hauman JH, Heersma H, Hide W, Beyers N, van Helden PD: Molecular evolution of Mycobacterium tuberculosis: phylogenetic reconstruction of clonal expansion. Tuberculosis (Edinb) 2001, 81:291-302.
• [20]Huong NT, Duong BD, Co NV, Quy HT, Tung LB, Bosman M, Gebhardt A, Velema JP, Broekmans JF, Borgdorff MW: Establishment and development of the National Tuberculosis Control Programme in Vietnam. Int J Tuberc Lung Dis 2005, 9:151-156.
• [21]Treatment of tuberculosis: Guidelines. 4th edition. WHO report; 2009. http://whqlibdoc.who.int/publications/2010/9789241547833_eng.pdf webcite, WHO/HTM/TB/2009.420
• [22]Menzies D, Benedetti A, Paydar A, Martin I, Royce S, Pai M, Vernon A, Lienhardt C, Burman W: Effect of duration and intermittency of rifampin on tuberculosis treatment outcomes: a systematic review and meta-analysis. PLoS Med 2009, 6:e1000146.
• [23]Quy HT, Cobelens FG, Lan NT, Buu TN, Lambregts CS, Borgdorff MW: Treatment outcomes by drug resistance and HIV status among tuberculosis patients in Ho Chi Minh City, Vietnam. Int J Tuberc Lung Dis 2006, 10:45-51.
• [24]Kiet VS, Lan NT, An DD, Dung NH, Hoa DV, van Vinh Chau N, Chinh NT, Farrar J, Caws M: Evaluation of the MTBDRsl test for detection of second-line-drug resistance in Mycobacterium tuberculosis. J Clin Microbiol 2010, 48:2934-2939.
• [25]Minh NN, Van Bac N, Son NT, Lien VT, Ha CH, Cuong NH, Mai CT, Le TH: Molecular characteristics of rifampin- and isoniazid-resistant Mycobacterium tuberculosis strains isolated in Vietnam. J Clin Microbiol 2012, 50:598-601.
• [26]Mathuria JP, Nath G, Samaria JK, Anupurba S: Molecular characterization of INH-resistant Mycobacterium tuberculosis isolates by PCR-RFLP and multiplex-PCR in North India. Infect Genet Evol 2009, 9:1352-1355.
• [27]Zhang M, Yue J, Yang YP, Zhang HM, Lei JQ, Jin RL, Zhang XL, Wang HH: Detection of mutations associated with isoniazid resistance in Mycobacterium tuberculosis isolates from China. J Clin Microbiol 2005, 43:5477-5482.
• [28]Piatek AS, Telenti A, Murray MR, El-Hajj H, Jacobs WR Jr, Kramer FR, Alland D: Genotypic analysis of Mycobacterium tuberculosis in two distinct populations using molecular beacons: implications for rapid susceptibility testing. Antimicrob Agents Chemother 2000, 44:103-110.
• [29]Moore DA, Shah NS: Alternative methods of diagnosing drug resistance–what can they do for me? J Infect Dis 2011, 204(Suppl 4):S1110-S1119.
• [30]Buu TN, van Soolingen D, Huyen MN, Lan NT, Quy HT, Tiemersma EW, Kremer K, Borgdorff MW, Cobelens FG: Increased transmission of Mycobacterium tuberculosis Beijing genotype strains associated with resistance to streptomycin: a population-based study. PLoS One 2012, 7:e42323.
• [31]van Soolingen D, Qian L, de Haas PE, Douglas JT, Traore H, Portaels F, Qing HZ, Enkhsaikan D, Nymadawa P, van Embden JD: Predominance of a single genotype of Mycobacterium tuberculosis in countries of east Asia. J Clin Microbiol 1995, 33:3234-3238.