Comparative Analysis of Levofloxacin Resistant Genes in Clinically Isolated Streptococcus pneumoniae

임상에서 분리한 Streptococcus pneumoniae에서 Levofloxacin 내성유전자의 비교 연구

  • Choi, Jae Min (Department of Biotechnology, Hannam University) ;
  • Park, Seon Hui (Department of Biotechnology, Hannam University) ;
  • Yoon, Ji A (Department of Biotechnology, Hannam University) ;
  • Han, Yang Keum (Department of Dental Hygiene, Daejeon Health Science College) ;
  • Lee, In Soo (Department of Biotechnology, Hannam University)
  • 최재민 (한남대학교 생명공학과) ;
  • 박선희 (한남대학교 생명공학과) ;
  • 윤지아 (한남대학교 생명공학과) ;
  • 한양금 (대전보건대학교 치위생(학)과) ;
  • 이인수 (한남대학교 생명공학과)
  • Published : 2012.04.30

Abstract

One hundred seventy four Streptococcus pneumoniae clinical isolates were categorized depending on the types of specimens, the age and the gender, respectively. All isolates were analyzed the characteristics of the multi-drug resistance including levofloxacin antibiotics. In the results of analysis depending on the type of samples, it had been confirmed that sputum was the main source of pneumonia infection because 156 of 174 strains (89.7%) were isolated in sputum samples. The opportunity for isolating the S. pneumoniae that had tolerance to levofloxacin was increased in over 51 age patients group compared with other age and male group. Eight strains of isolates were evaluated higher resistant to levofloxacin, and those also showed multi-drug resistant including penicillin, tetracycline, erythromycin, clindamycin and trimethoprim-sulfamethoxazole. In the results of sequence analysis of quinolone resistance determining region in SP32 (MIC $64{\mu}g/mL$) and SP96 (MIC $8{\mu}g/mL$) which were levofloxacin resistant strains, an amino acid substitutions were found Ser-81$\rightarrow$Phe in both GyrA of SP32 and SP96, and Ser-11$\rightarrow$Gly in only SP96. A Ser-79$\rightarrow$Phe substitution of ParC was found in both.

임상에서 분리한 총 174 균주의 S. pneumoniae를 대상으로 검체별, 연령별, 성별 분리빈도 및 levofloxacin의 내성도를 조사하였으며, 항생제 감수성 검사를 통해 다제내성도를 확인하였다. S. pneumoniae가 가장 많이 분리된 검체는 객담으로서 총 174 균주의 89.7%인 156 균주가 분리되었다. 특히 남성과 51세의 고령환자에서 분리빈도가 높았으며, 분리된 levofloxacin 내성 8 균주 모두는 penicillin, tetracycle, erytromycin, clindamycin 및 trimethoprim-sulfamethoxazoe 대한 다제내성도 함께 소유하고 있는 것으로 확인되었다. 분리된 levofloxacin 내성균주 SP32 (MIC $64{\mu}g/mL$)와 SP96 (MIC $8{\mu}g/mL$)의 QRDR 염기서열을 분석한 결과, SP32와 SP96 균주의 GyrA에서 Ser-81$\rightarrow$Phe로, SP96에서 Ser-11$\rightarrow$Gly으로 아미노산 치환이 각각 확인되었고, ParC에서는 두 균주 모두 Ser-79$\rightarrow$Phe으로 치환된 돌연변이가 확인되었다.

Keywords

References

  1. Kim KH et al.: The causative organism of bacterial meningitis in Korean children, 1986-1995. J Korean Med Sci 13(1): 60-64, 1998.
  2. Kaplan VD et al.: Hospitalized community-acquired pneumonia in the elderly: age-and sex-related patterns of care and outcome in the United States. Am J Respir Crit Care Med 165(6): 766-772, 2002. https://doi.org/10.1164/ajrccm.165.6.2103038
  3. Janssens J P: Pneumonia in the elderly (geriatiric) population. Curr Opin Pulm Med 11(3): 226-30, 2005.
  4. 최원석 등: 국내 침습성 폐렴사슬알균 감염증의 임상양상. Infection & Chemotherapy 42(3): 156-161, 2010. https://doi.org/10.3947/ic.2010.42.3.156
  5. Gutierrez FM et al.: The influence of age and gender on the population-based incidence of community-acquired pneumonia caused by different microbial pathogens. J Infect 53(3): 166-174, 2006. https://doi.org/10.1016/j.jinf.2005.11.006
  6. Klugman KP et al.: Drug resistance patterns and serogroups or serotypes of pneumococcal isolates from cerebrospinal fluid or blood. J Infect Dis 158(5): 956-964, 1988. https://doi.org/10.1093/infdis/158.5.956
  7. Marisa Haenni et al.: Mutational Analysis of Class A and Class B Penicillin-Binding Proteins in Streptococcus gordonii. Antimicrob. Aqents and Chemother. 50(12): 4062-4069, 2006. https://doi.org/10.1128/AAC.00677-06
  8. Chen DK et al.: Decreased susceptibility of Streptococcus pneumoniae to fluoroquinolones in Canada. N Engl J Med 341(4): 233-239, 1999. https://doi.org/10.1056/NEJM199907223410403
  9. Doern GV et al.: Antimicrobial resistance among clinical isolates of Streptococcus pneumoniae in the United states during 1999-2000, including a comparison of resistance rates since1994-1995. Antimicrob Agents Chemother 45(6): 1721-1729, 2001. https://doi.org/10.1128/AAC.45.6.1721-1729.2001
  10. Gonzalez Irene M et al.: Fluoroquinolone resistance mutations in the parC, parE, and gyrA genes of clinical isolates of viridans group Streptococci. Antimicrob Aqents Chemother 42(11): 2792-2798, 1998.
  11. Janoir C et al.: High-level fluoroquinolone resistance in Streptococcus pneumoniae requires mutations in parC and gyrA. Antimicrob Aqents Chemothe 40(12): 2760-2764, 1996.
  12. Yoshida H et al.: Quinolone-resistant mutations of the gyrA gene of Escherichia coli. Mol Gen Genet 211(1): 1-7, 1988. https://doi.org/10.1007/BF00338386
  13. Balsalobre L. et al.: Viridans group streptococci are donors in horizental transfer of topoisomerase IV genes to Streptoccus pneumoniae. Antimicrob. Agents and Chemother. 47(12): 2072-2081. 2003.
  14. Margaret Ip et al.: Fluoroquinolone Resistance in Atypical Pneumococci and Oral Streptococci: Evidence of Horizontal Gene Transfer of Fluoroquinolone Resistance Determinants from Streptococcus pneumoniae. Antimicrob. Agents and Chemother. 51(8): 2690-2700, 2007. https://doi.org/10.1128/AAC.00258-07
  15. 송재훈 등: 지역사회획득 폐렴의 치료지침 권고안. Tuberc Respir Dis 67(4): 281-302, 2009. https://doi.org/10.4046/trd.2009.67.4.281
  16. Bowen MK et al.: The Optochin sensitivity test: a reliable method for identification of pneumococci. J Lab Clin Med 49(4): 641-642, 1957.
  17. MacFaddin JF: Biochemical tests for identification of medical bacteria(1st). The williams & wilkins Co., Baltimore pp. 9-15, 1976.
  18. Clinical and Laboratory Standards Institute: Performance standards for antimicrobial susceptibility testing; Nineteenth informational supplement. CLSI document M100-S19, CLSI, 2009.
  19. Kim HS et al.:. Evaluation of microscan microSTREP plus antimicrobial suscepttibility panel for testing Streptococcus pneumoniae. Korean J Clin Microbiol 11(1): 18-22, 2008. https://doi.org/10.5145/KJCM.2008.11.1.18
  20. Clinical and Laboratory Standards Institute. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard-seventh edition. CLSI document M7-A7. Wayne, PA; CLSI, 2006.
  21. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing; sixteenth informational supplement. CLSI document M100-S16. Wayne, PA; CLSI, 2006.
  22. Jeffrey H, Miller: Methods in enzymology. Bacterial Genetic Systems, pp. 72-74, 1991.
  23. Tleyjeh IM et al.: The impact of penicillin resistance on short-term mortality in hospitalized adults with Pneumococcal pneumonia: a systematic review and meta-analysis. Clin Infect Dis 42(6): 788-797, 2006. https://doi.org/10.1086/500140
  24. Pestova E et al.: Intracellular targets of moxi-floxacin: a comparison with other fluoroquinolones. J Antimicrob Chemother 45(5): 583-590, 2000. https://doi.org/10.1093/jac/45.5.583