بررسی فراوانی ژن های مقاومت کینولونی وابسته به پلاسمید در جدایه های بالینی اشریشیا کلی و گونه های کلبسیلا در تهران
محورهای موضوعی : میکروب شناسی پزشکیمعصومه محمدبیگی 1 , جعفر اکبر مهر 2 , بهبود جعفری 3
1 - کارشناس ارشد، گروه میکروب شناسی، واحد اهر، دانشگاه آزاد اسلامی، اهر
2 - استادیار، گروه میکروب شناسی، واحد سراب،
دانشگاه آزاد اسلامی، سراب
3 - استادیار، گروه میکروب شناسی، واحد اهر، دانشگاه آزاد اسلامی، اهر
کلید واژه: اشریشیا کلی, کلبسیلا, ژن های qnr, ژن aac(6)-Ib-cr,
چکیده مقاله :
سابقه و هدف: حضور ژن های پلاسمیدی qnr در باکتری ها عامل اصلی ایجاد مقاومت در برابر کینولون ها به شمار می آید. این مطالعه با هدف ارزیابی حضور ژن های qnr وابسته به پلاسمید و ژن های مقاوم aac(6)-Ib-cr در جدایه های بالینی اشریشیاکلی و کلبسیلا جداسازی شده از تهران انجام شد. مواد و روش ها: در این مطالعه مقطعی تعداد 132 جدایه اشریشیاکلی و 98 جدایه کلبسیلا از بیماران مبتلا به عفونت ادراری مراجعه کننده به بیمارستان آیت الله طالقانی تهران جمع آوری گردید. مقاومت آنتی بیوتیکی با استفاده از روش کربی-بائر و معیار CLSI انجام شد. پس از استخراج ژنوم، به منظور تکثیر ژن های qnr و aac(6)-Ib-cr از روش واکنش زنجیره ای پلی مراز (PCR) استفاده گردید. یافته ها: در مجموع 47.72 درصد از جدایه های اشریشیا کلی نسبت به آنتی بیوتیک سیپروفلوکساسین و 62.12 درصد نسبت به نالیدیکسیک اسید مقاوم بودند. همچنین 38.77 درصد از جدایه های کلبسیلا در برابر سیپروفلوکساسین و 51.02 درصد نسبت به نالیدیکسیک اسید مقاوم بودند. در مطالعه حاضر شیوع ژن های aac(6)-lb-cr، qnrA، qnrB وqnrS در جدایه های اشریشیا کلی به ترتیب 30.18، 37.73، 67.92 و 33.96 درصد و در کلبسیلا به ترتیب 31.42، 22.85، 68.57 و 31.42 درصد گزارش شد. نتیجه گیری: نتایج این مطالعه نشان دهنده فراوانی نسبتاً بالای ژن های qnr در جدایه های اشریشیاکلی و کلبسیلا مقاوم به سیپروفلوکساسین در شهر تهران بود.
Background & Objectives: The qnr gene is a plasmid carrying resistant genes involved in bacterial resistance to quinolone resistance plasmid factors. The aim of this study was to evaluate the presence of either gene qnr, a plasmid resistant gene, and aac (6) -Ib-cr, from Escherichia coli and Klebsiella spp isolated from clinical samples. Materials & Methods: In this sectional study, 132 E. coli and 98 Klebsiella isolates were collected from patients with the UTI referred to the ayatollah Taleghani Hospital in Tehran. Antibiotic resistance was evaluated using Kirby-Bauer and CLSI criteria. After DNA extraction, qnr and aac(6)-Ib-cr genes were amplified using PCR method. Results: In this study, E. coli strains were resistant to ciprofloxacin and nalidixic acid in a ratio of 47.72 % and 62.12 %, respectively. Also, Klebsiella isolates were resistant to ciprofloxacin and nalidixic acid in a ratio of 38.77 % and 51.02 %, respectively. In the present study, the frequency of aac(6)-lb-cr, qnrA , qnrB, qnrS genes in E. coli were 30.18, 37.73, 67.92, 33.96 %. Also these frequency were 31.42, 22.58, 68.57, 31.42 % in Klebsiella. Conclusion: This study showed high frequency of the ciprofloxacin resistant qnr gense in E. coli and Klebsiella isolates from Tehran.
1. Van den Bogaard AE, Stobberingh EE. Epidemiology of resistance to antibiotics: links between animals and humans. Int J Antimicrob Agent. 2000; 14(4): 327-335.
2. Wada K, Kariyama R, Mitsuhata R, Uehara S, Watanabe T, Monden K. Experimental and clinical studies on fluroquinolone- insusceptible Escherichia coli isolated from patient with urinary tract infections from 1994-2007. Acta Med Okayama. 2009; 63(5): 263-272.
3. Nazik H, Bektore B, Ongen B, Ozuyrt M, Baylan O, Haznedaroglu T. Co-expression of plasmid-mediated quinolone resistance-qnrA1 and blaVEB1 gene in Providensia staturii strain. New Microbiol. 2011; 34(2): 225-228.
4. Robicsek A, Jacoby GA, Hooper DC. The worldwide, emergence of plasmid mediated quinolone resistance. Lancet Infect Dis. 2006; 6(1): 629-640.
5. Zhou TL, Chen XJ, Zhou MM, Zhao YJ, Luo HZ, Bao QU. Prevalence of plasmid-mediated quinolone resistance in Escherichia coli isolated in Wenzhou, southern China, 2002-2008. Jpn J Infect Dis. 2011; 64(1): 55-57.
6. Cattoir V, Poirel L, Rotimi V, Soussy CJ, Nordman P. Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing entrobacterial isolated. J Antimicrob Chemother. 2007; 60(4): 394-397.
7. Corkill JE, Anson JJ, Hart CA. High prevalence of the plasmid-mediated quinolone resistance determinant qnrA in multidrug-resistant Enterobacteriaceae from blood cultures in Liverpool, UK. J Antimicrob Chemother. 2005; 56(6): 1115-1117.
8. Rodríguez-Martínez JM, Cano ME, Velasco C, Martínez-Martínez L, Pascual Á. Plasmid-mediated quinolone resistance: an update. J Infection Chemotherapy. 2011; 17(2): 149-182.
9. DiPersio JR, Deshpande LM, Biedenbach DJ, Toleman MA, Walsh TR, Jones RN. Evolution and dissemination of extended-spectrum β-lactamase-producing Klebsiella pneumoniae: Epidemiology and molecular report from the sentry antimicrobial surveillance program (1997–2003). Diagn Microb Infect Dis. 2005; 51(1): 1-7.
10. Kim HB, Park CH, Kim CJ, Kim E-C, Jacoby GA, Hooper DC. Prevalence of plasmid- mediated quinolone resistance determinants over a 9-year period. Antimicrob Agents Chemotherapy. 2009; 53(2): 639-645.
11. Akbari-Nakhjavani F, Mirsalehi A, Hamidian M, Kazemi B, Mirafshar M, Jabal Ameli F. Antimicrobial susceptibility testing of Escherichia coli strains isolated from urinary tract infections to fluoroquinolones and detection of gyrA mutations in resistant strains. Daru. 2007; 15(2): 94-99.
12. Syeda QA, Ale Z, Baquar SN, Shahjahan Sh, Rabia B. Resistance pattern of ciprofloxacin against different pathogens. Oman Med J. 2010; 25(4): 294.
13. Colodner R, Kometiani I, Chazan B, Raz R. Risk factors for community-acquired urinary tract infection due to quinolone-resistant E. coli. Infection. 2008; 36(1): 41-45.
14. Santiso R, Tamayo M, Fernández JL, del Carmen Fernández M, Molina F, Villanueva R. Rapid and simple determination of ciprofloxacin resistance in clinical strains of Escherichia coli. J Clin Microbiol. 2009; 47(8): 2593-2595.
15. Molla Zadeh H, Sheikh Haji Zadeh B, Aslamyk, Hamidi M, Bahmanabadi R. Susceptibility patterns of antibiotic resistance in K. pneumoniae strains isolated from clinical samples Arad hospital in Tehran. Iran J Infectious Dis Trop Med J. 2013; 18(62): 37-41.
16. Zibaei M, Firoozeh F. Detection of qnrA gene among quinolone-resistant Escherichia coli isolated from urinary tract infections in Khorram Abad during 2011-2012. Kashan Uni Med Sci J. 2013;17(5): 488-494.
17. Paterson DL, Bonomo RA. Extended-spectrum β-lactamases: a clinical update. Clin Microbiol Rev. 2005; 18(4): 657-686.
18. Clinical and laboratory standard institute. Performance standard for antimicrobial susceptibility testing. M100-S25: CLSI; 2015.
19. Hokmi vala M, Abdi Sh, Bagheri bejestani F. QnrA gene in fluoroquinolone-resistant Escherichia coli isolates scattering isolated from the urine of patients referred to Imam Khomeini Tehran. J Infect Dis. 2014; 19(64): 63-66. [In Persian]
20. Mansouri Jamshidi N1, Pakzad I, Tabaraei B, Hadadi A. Evaluating the frequency of ciprofloxacin resistance qnr genes in Escherichia coli strains isolated from clinical samples of Imam Khomani and Milad Hospitals in Ilam and Tehran, Iran. J Isfahan Univ Med Sci. 2013; 21(6): 16-22.[In Persian]
21. Nkat G, Muller G, Braissant O, Frei R, Tschudin-Suter S, Rieken M. Increasing prevalence of ciprofloxacin resistance in extended spectrum betalactamase-producing Escherichia coli urinary isolates. World J Urol. 2013; 31: 1427-1432.
22. Nsal S, Tandon V. Contribution of mutations in DNA gyrase and topoisomerase IV genes to ciprofloxacin resistance in Escherichia coli clinical isolates. Int J Antimicrob Agent. 2011; 37: 253-255.
23. In JH, Jung HJ, Lee JY, Kim HR, Lee JN, Chang CL. High rates of plasmid-mediated quinolone resistance qnrB variants among ciprofloxacin-resistant Escherichia coli and K. pneumoniae from urinary tract infections in Korea. Microb Drug Resist. 2008; 14: 221-226.
24. Mirmostafa S, haddadi A, Mozaffari Sabet3 N. Determination of plasmid profile and antibiotic resistance pattern in clinical Escherichia coli strains, isolated from different region of Karaj city. New Cell Mol Biotechnol J. 2014; 3(12): 93-97.
25. Oktem IMA, Gulay Z, Biçmen M, Gur D, Group HPS. qnrA prevalence in extended-spectrum beta-lactamase-positive Enterobacteriaceae isolates from Turkey. Jpn J Infect Dis. 2008; 61(1): 13.
26. Karah N, Poirel L, Bengtsson S, Sundqvist M, Kahlmeter G, Nordmann P. Plasmid-mediated quinolone resistance determinants qnr and aac (6′)-Ib-cr in Escherichia coli and Klebsiella spp. from Norway and Sweden. Diagn Microbiol Infect Dis. 2010; 66(4): 425-431.
27. Zhou T-L, Chen X-J, Zhou M-M, Zhao Y-J, Luo X-H, Bao Q-Y. Prevalence of plasmid- mediated quinolone resistance in Escherichia coli isolates in Wenzhou, Southern China, 2002-2008. Jpn J Infect Dis. 2011; 64(1): 55-57.
28. Azadpour M, Soleimani Y, Rezaie F, Nikanpour E, Mahmoudvand H, Jahanbakhsh S. Prevalence of qnr genes and antibiotic susceptibility patterns among clinical isolates of Klebsiella pneumoniae in West of Iran. J Bacteriol Parasitol. 2014; 5(5): 1. [In Persian]
29. Shin SY, Kwon KC, Park JW, Song JH, Ko YH, Sung JY. Characteristics of aac (6')-Ib-cr gene in extended-spectrum β-lactamase-producing Escherichia coli and K. pneumoniae isolated from Chungnam area. Korean J Lab Med. 2009; 29(6): 541-550.
_||_1. Van den Bogaard AE, Stobberingh EE. Epidemiology of resistance to antibiotics: links between animals and humans. Int J Antimicrob Agent. 2000; 14(4): 327-335.
2. Wada K, Kariyama R, Mitsuhata R, Uehara S, Watanabe T, Monden K. Experimental and clinical studies on fluroquinolone- insusceptible Escherichia coli isolated from patient with urinary tract infections from 1994-2007. Acta Med Okayama. 2009; 63(5): 263-272.
3. Nazik H, Bektore B, Ongen B, Ozuyrt M, Baylan O, Haznedaroglu T. Co-expression of plasmid-mediated quinolone resistance-qnrA1 and blaVEB1 gene in Providensia staturii strain. New Microbiol. 2011; 34(2): 225-228.
4. Robicsek A, Jacoby GA, Hooper DC. The worldwide, emergence of plasmid mediated quinolone resistance. Lancet Infect Dis. 2006; 6(1): 629-640.
5. Zhou TL, Chen XJ, Zhou MM, Zhao YJ, Luo HZ, Bao QU. Prevalence of plasmid-mediated quinolone resistance in Escherichia coli isolated in Wenzhou, southern China, 2002-2008. Jpn J Infect Dis. 2011; 64(1): 55-57.
6. Cattoir V, Poirel L, Rotimi V, Soussy CJ, Nordman P. Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing entrobacterial isolated. J Antimicrob Chemother. 2007; 60(4): 394-397.
7. Corkill JE, Anson JJ, Hart CA. High prevalence of the plasmid-mediated quinolone resistance determinant qnrA in multidrug-resistant Enterobacteriaceae from blood cultures in Liverpool, UK. J Antimicrob Chemother. 2005; 56(6): 1115-1117.
8. Rodríguez-Martínez JM, Cano ME, Velasco C, Martínez-Martínez L, Pascual Á. Plasmid-mediated quinolone resistance: an update. J Infection Chemotherapy. 2011; 17(2): 149-182.
9. DiPersio JR, Deshpande LM, Biedenbach DJ, Toleman MA, Walsh TR, Jones RN. Evolution and dissemination of extended-spectrum β-lactamase-producing Klebsiella pneumoniae: Epidemiology and molecular report from the sentry antimicrobial surveillance program (1997–2003). Diagn Microb Infect Dis. 2005; 51(1): 1-7.
10. Kim HB, Park CH, Kim CJ, Kim E-C, Jacoby GA, Hooper DC. Prevalence of plasmid- mediated quinolone resistance determinants over a 9-year period. Antimicrob Agents Chemotherapy. 2009; 53(2): 639-645.
11. Akbari-Nakhjavani F, Mirsalehi A, Hamidian M, Kazemi B, Mirafshar M, Jabal Ameli F. Antimicrobial susceptibility testing of Escherichia coli strains isolated from urinary tract infections to fluoroquinolones and detection of gyrA mutations in resistant strains. Daru. 2007; 15(2): 94-99.
12. Syeda QA, Ale Z, Baquar SN, Shahjahan Sh, Rabia B. Resistance pattern of ciprofloxacin against different pathogens. Oman Med J. 2010; 25(4): 294.
13. Colodner R, Kometiani I, Chazan B, Raz R. Risk factors for community-acquired urinary tract infection due to quinolone-resistant E. coli. Infection. 2008; 36(1): 41-45.
14. Santiso R, Tamayo M, Fernández JL, del Carmen Fernández M, Molina F, Villanueva R. Rapid and simple determination of ciprofloxacin resistance in clinical strains of Escherichia coli. J Clin Microbiol. 2009; 47(8): 2593-2595.
15. Molla Zadeh H, Sheikh Haji Zadeh B, Aslamyk, Hamidi M, Bahmanabadi R. Susceptibility patterns of antibiotic resistance in K. pneumoniae strains isolated from clinical samples Arad hospital in Tehran. Iran J Infectious Dis Trop Med J. 2013; 18(62): 37-41.
16. Zibaei M, Firoozeh F. Detection of qnrA gene among quinolone-resistant Escherichia coli isolated from urinary tract infections in Khorram Abad during 2011-2012. Kashan Uni Med Sci J. 2013;17(5): 488-494.
17. Paterson DL, Bonomo RA. Extended-spectrum β-lactamases: a clinical update. Clin Microbiol Rev. 2005; 18(4): 657-686.
18. Clinical and laboratory standard institute. Performance standard for antimicrobial susceptibility testing. M100-S25: CLSI; 2015.
19. Hokmi vala M, Abdi Sh, Bagheri bejestani F. QnrA gene in fluoroquinolone-resistant Escherichia coli isolates scattering isolated from the urine of patients referred to Imam Khomeini Tehran. J Infect Dis. 2014; 19(64): 63-66. [In Persian]
20. Mansouri Jamshidi N1, Pakzad I, Tabaraei B, Hadadi A. Evaluating the frequency of ciprofloxacin resistance qnr genes in Escherichia coli strains isolated from clinical samples of Imam Khomani and Milad Hospitals in Ilam and Tehran, Iran. J Isfahan Univ Med Sci. 2013; 21(6): 16-22.[In Persian]
21. Nkat G, Muller G, Braissant O, Frei R, Tschudin-Suter S, Rieken M. Increasing prevalence of ciprofloxacin resistance in extended spectrum betalactamase-producing Escherichia coli urinary isolates. World J Urol. 2013; 31: 1427-1432.
22. Nsal S, Tandon V. Contribution of mutations in DNA gyrase and topoisomerase IV genes to ciprofloxacin resistance in Escherichia coli clinical isolates. Int J Antimicrob Agent. 2011; 37: 253-255.
23. In JH, Jung HJ, Lee JY, Kim HR, Lee JN, Chang CL. High rates of plasmid-mediated quinolone resistance qnrB variants among ciprofloxacin-resistant Escherichia coli and K. pneumoniae from urinary tract infections in Korea. Microb Drug Resist. 2008; 14: 221-226.
24. Mirmostafa S, haddadi A, Mozaffari Sabet3 N. Determination of plasmid profile and antibiotic resistance pattern in clinical Escherichia coli strains, isolated from different region of Karaj city. New Cell Mol Biotechnol J. 2014; 3(12): 93-97.
25. Oktem IMA, Gulay Z, Biçmen M, Gur D, Group HPS. qnrA prevalence in extended-spectrum beta-lactamase-positive Enterobacteriaceae isolates from Turkey. Jpn J Infect Dis. 2008; 61(1): 13.
26. Karah N, Poirel L, Bengtsson S, Sundqvist M, Kahlmeter G, Nordmann P. Plasmid-mediated quinolone resistance determinants qnr and aac (6′)-Ib-cr in Escherichia coli and Klebsiella spp. from Norway and Sweden. Diagn Microbiol Infect Dis. 2010; 66(4): 425-431.
27. Zhou T-L, Chen X-J, Zhou M-M, Zhao Y-J, Luo X-H, Bao Q-Y. Prevalence of plasmid- mediated quinolone resistance in Escherichia coli isolates in Wenzhou, Southern China, 2002-2008. Jpn J Infect Dis. 2011; 64(1): 55-57.
28. Azadpour M, Soleimani Y, Rezaie F, Nikanpour E, Mahmoudvand H, Jahanbakhsh S. Prevalence of qnr genes and antibiotic susceptibility patterns among clinical isolates of Klebsiella pneumoniae in West of Iran. J Bacteriol Parasitol. 2014; 5(5): 1. [In Persian]
29. Shin SY, Kwon KC, Park JW, Song JH, Ko YH, Sung JY. Characteristics of aac (6')-Ib-cr gene in extended-spectrum β-lactamase-producing Escherichia coli and K. pneumoniae isolated from Chungnam area. Korean J Lab Med. 2009; 29(6): 541-550.