Tuberculosis and Respiratory Diseases

The Korean Academy of Tuberculosis and Respiratory Diseases (대한결핵및호흡기학회)
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Cross Resistance of Fluoroquinolone Drugs on gyrA Gene Mutation in Mycobacterium tuberculosis

결핵균에서 gyrA 유전자 돌연변이에 따른 Fluoroquinolone계 약제들의 교차내성

  • Park, Young Kil (Korean Institute of Tuberculosis) ;
  • Park, Chan Hong (Korean Institute of Tuberculosis) ;
  • Koh, Won-Jung (Division of Pulmonary and Critical Care Medicine, Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine) ;
  • Kwon, O Jung (Division of Pulmonary and Critical Care Medicine, Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine) ;
  • Kim, Bum Jun (Department of Microbiology, Seoul National University College of Medicine) ;
  • Kook, Yoon Hoh (Department of Microbiology, Seoul National University College of Medicine) ;
  • Cho, Sang Nae (Department of Microbiology, Yonsei University College of Medicine) ;
  • Chang, Chulhun (Korean Institute of Tuberculosis) ;
  • Bai, Gill Han (Korean Institute of Tuberculosis)
  • 박영길 (대한결핵협회 결핵연구원) ;
  • 박찬홍 (대한결핵협회 결핵연구원) ;
  • 고원중 (성균관대학교 의과대학 내과학교실 삼성서울병원 호흡기내과) ;
  • 권오정 (성균관대학교 의과대학 내과학교실 삼성서울병원 호흡기내과) ;
  • 김범준 (서울대학교 의과대학 미생물학교실) ;
  • 국윤호 (서울대학교 의과대학 미생물학교실) ;
  • 조상래 (연세대학교 의과대학 미생물학교실) ;
  • 장철훈 (대한결핵협회 결핵연구원) ;
  • 배길한 (대한결핵협회 결핵연구원)
  • Received : 2005.06.09
  • Accepted : 2005.08.16
  • Published : 2005.09.30
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Abstract

Background : Fluoroquinolone drugs are an important anti-tuberculous agent for the treatment of multi-drug resistant tuberculosis. However, many drugs belonging to the fluoroquinolones have different cross resistance to each other. Methods : Sixty-three ofloxacin (OFX) resistant and 10 pan-susceptible M. tuberculosis isolates were selected, and compared for their cross resistance using a proportion method on Lowenstein-Jensen media, containing ofloxacin (OFX), ciprofloxacin (CIP), levofloxacin (LVX), moxifloxacin (MXF), gatifloxacin (GAT) and sparfloxacin (SPX), at concentrations ranging from 0.5 to $3{\mu}g/ml$. DNA extracted from the isolates was directly sequenced after amplifying from the gyrA and gyrB genes. Results : The 63 OFX resistant M. tuberculosis isolates showed complete cross resistance to CIP, but only 90.5, 44.4, 36.5 and 46.0% to LVX, MXF, GAT, and to SPX, respectively. Fifty-one of the isolates (81.0%) had point mutations in codons 88, 90, 91 and 94 in gyrA, which are known to be correlated with OFX resistance. The Gly88Ala, Ala90Valand Asp94Ala mutations in gyrA showed a tendency to be susceptible to MXF, GAT and SPX. Only 4 isolates had mutations in the gyrB gene, which did not affect the OFX resistance. Conclusion : About 60% of the OFX resistant M. tuberculosis isolates were susceptible to GAT, SPX and MXF. These fluoroquinolones may be useful in the treatment of TB patients showing OFX resistance.

배 경 : FQ계 약제는 다제내성 결핵환자들의 치료를 위한 주용 2차 항결핵약제의 하나이다. 그러나 FQ계 여러 약제들 간 교차내성정도와 내성관련 유전자인 gyrA의 돌연변이간 관련성을 조사하게 되었다. 방 법 : 결핵균 중에서 OFX 내성인 63균주와 감수성인 10균주를 대상으로 Lowenstein-Jensen 배지에 CIP, LVX, MXF, GAT, SPX 등에 대한 교차내성을 조사하였다. 퀴놀론계 각 약제를 $0.5-3{\mu}g/ml$ 농도로 첨가한 배지에서, $2{\mu}g/ml$ 이상에서 균이 발육한 경우를 내성으로 판정하였다. OFX 내성인 균주의 gyrA 및 gyrB 유전자의 돌연변이가 잘 발생하는 곳을 염기서열 분석하여 약제별로 그 교차내성 양상을 비교하였다. 결 과 : OFX 내성인 63균주 모두가 CIP약제에서는 교차내성을 나타내었고, LVX에는 90.5%, MXF에는 44.4%, GAT에는 36.5%, SPX에는 46.0%가 교차내성을 보였다. 이들 내성균주의 81.0%가 gyrA 유전자에 돌연변이를 가지고 있었으며, 그 분포는 아미노산 88번, 90번, 91번, 94번에서 나타났다. 그 중에서도 Gly88Ala, Ala90Val, Asp94Ala 돌연변이는 제3세대 FQ계 약제인 MXF, GAT, SPX에 감수성을 나타내는 경향이 있었다. 한편 gyrB 유전자에서는 63개 OFX 내성균주 중 4균주만이 돌연변이를 나타내었으나, 이들과 gyrB 유전자내 돌연변이가 없는 균주들 사이에서 FQ의 내성에 관련해서 차이가 없었다. 결 론 : OFX 내성균 중에서 약 60% 정도가 제3세대 FQ계 약제에서 감수성을 보였다. 따라서 이 들 약제에 대해서는 별도로 감수성검사를 실시하여 처방에 활용한다면, OFX 내성환자의 치료를 크게 향상시킬 수 있을 것으로 판단되었다.

Keywords

References

  1. Cambau E, Sougakoff W, Besson M, Truffot-Pernot C, Grosset J, Jarlier V. Selection of a gyrA mutant of Mycobacterium tuberculosis resistant to fluoroquinꠀ olones during treatment with ofloxacin. J Infect Dis 1994;170:479-83
  2. Grosset JH. Treatment of tubeculosis in HIV infection. Tuberc Lung Dis 1992;73:378-83 https://doi.org/10.1016/0962-8479(92)90044-K
  3. Tsukamura M, Nakamura E, Yoshii S, Amano H. Therapeutic effect of a new antibacterial substance ofloxacin (DL8280) on pulmonary tuberculosis. Am Rev Respir Dis 1985;131:352-6
  4. Blumberg HM, Burman WJ, Chaisson RE, Daley CL, Etkind SC, Friedman LN, et al. American Thoracis Society/Centers for Disease Control and Prevention/ Infectious Diseases Society of America: treatment of tuberculosis. Am J Respir Crit Care Med 2003;167: 603-62 https://doi.org/10.1164/rccm.167.4.603
  5. Crofton J, Chaulet P, Maher D, Grosset J, Harris W, Horne N, Iseman M, Watt B. Guidelines for the management of drug-resistant tuberculosis. Geneva: World Health Organization; 1997
  6. Casal M, Ruiz P, Herreras A. Study of the in vitro susceptibility of M. tuberculosis to ofloxacin in Spain. Int J Tuberc Lung Dis 2000;4:588-91
  7. Hoffner SF, Gezelius I, Olsson-Liljequist B. In vitro activity of fluorinated quinolones and macrolides against drug-resistant Mycobacterium tuberculosis. J Antimicrob Chemother 1997;40:885-8 https://doi.org/10.1093/jac/40.6.885
  8. Yu MC, Suo J, Lin TP, Luh KT. In vitro activity of ofloxacin against Mycobacterium tuberculosis. J Formos Med Assoc 1997;96:13-6
  9. Takiff H, Salazar L, Guerrero C, Philipp W, Huang WM, Kreiswirth B, et al. Cloning and nucleotide seꠀ quence of the Mycobacterium tuberculosis gyrA and gyrB genes, and characterization of quinolone resistance mutations. Antimicrob Agents Chemother 1994; 38:773-80
  10. Kim SJ, Bai GH, Hong YP. Drug resistant tuberꠀ culosis in Korea, 1994. Int J Tuberc Lung Dis 1997;1: 302-8
  11. Sullivan EA, Kreiswirth BN, Palumbo L, Kapur V, Musser JM, Ebrahimzadeh A, et al. Emergence of fluoroquinolone-resistant tuberculosis in New York City. Lancet 1995;345:1148-50 https://doi.org/10.1016/S0140-6736(95)90980-X
  12. Dong Y, Xu C, Zhao X, Domagala J, Drlica K. Fluoꠀ roquinolne action against mycobacteria: effects of C-8 substituents on growth, survival, and resistance. Antimicrob Agents Chemother 1998;42:2978-84
  13. Xu C, Kreiswirth BN, Sreevatsan S, Musser JM, Drlica K. Fluoroquinolone resistance associated with specific gyrase mutations in clinical isolates of multidrug resistant Mycobacterium tuberculosis. J Infect Dis 1996; 174:1127-30
  14. Dong Y, Zhao X, Kreiswirth BN, Drlica K. Mutant prevention concentration as a measure of antibiotic potency: studies with clinical isolates of Mycobacterium tuberculosis. Antimcirob Agents Chemother 2000; 44:2581-4 https://doi.org/10.1128/AAC.44.9.2581-2584.2000
  15. Zhao BY, Pine R, Domagala J, Drlica K. Fluoroquiꠀ nolone action against clinical isolates of Mycobacterꠀ ium tuberculosis: effects of a C8-methoxyl group on survival in liquid media and in human macrophages. Antimicrob Agents Chemother 1999;43:661-6
  16. Drlica K, Zhao X. DNA gyrase, topoisomerase IV, and the 4-quinolones. Microbiol Mol Biol Rev 1997;61: 377-92
  17. Hiasa H, Yousef DO, Marians KJ. DNA strand cleavage is required for replication fork arrest by a frozen topoisomerase-quinolone DNA ternary complex. J Biol Chem 1996;271:26424-9 https://doi.org/10.1074/jbc.271.42.26424
  18. Manes SH, Pruss GJ, Drlica K. Inhibition of RNA synthesis by oxolinic acid is unrelated to average DNA supercoiling. J Bacteriol 1983;155:420-3
  19. Cheng AF, Yew WW, Chan EW, Chin ML, Hui MM, Chan RC. Multiplex PCR amplimer conformation analysis for rapid detection of gyrA mutations in fluoroquinolone-resistant Mycobacterium tuberculosis clinical isolates. Antimicrob Agents Chemother 2004;48: 596-601 https://doi.org/10.1128/AAC.48.2.596-601.2004
  20. Perlman DC, el Sadr WM, Heifets LB, Nelson ET, Matts JP, Chirgwin K, et al. Susceptibility to levofloxacin of Mycobacterium tuberculosis isolates from patients with HIV-related tuberculosis and characterization of a strain with levofloxacin monoresistacne. AIDS 1997;11:1473-8
  21. Kim BJ, Kang YS, Park SK. Activity of moxifloxacin against ofloxacin-resistant Mycobacterium tuberculosis: a study of cross-resistance between ofloxacin and moxifloxacin. Tuberc Respir Dis 2004;57:405-10
  22. Ji B, Lounis N, Maslo C, Truffot-Pernot C, Bonnafous P, Grosset J. In vitro and in vivo activities of moxifloxacin and clinafloxacin against Mycobacterium tuberculosis. Antimicrob Agents Chemother 1998;42: 2066-9
  23. Alvirez-Freites E, Carter J, Cynamon MH. In vitro and in vivo activities of gatifloxacin against Mycobacterium tuberculosis. Antimicrob Agents Chemother 2002;46:1022-5 https://doi.org/10.1128/AAC.46.4.1022-1025.2002
  24. Ji B, Lounis N, Truffot-Pernoc C, Grosset J. In vitro and in vivo activities of levofloxacin against Mycobacterium tuberculosis. Antimicrob Agents Chemother 1995;39:1341-4
  25. Canetti G, Fox W, Khomenko A, Mahler H, Menon N, Mitchison DA, et al. Advances in techniques of testing mycobacterial drug sensitivity and the use of sensitivity tests in tuberculosis control programmes. Bull Worl Health Organ 1969;41:21-43