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Manuscript ID : 686028 Visit : 235 Page: 419 - 429

10.22034/jchr.2021.686028

Article Type: Original Research

Detection of Gyrase Enzyme among Clinical Isolates of E. coli Resistance to some Quinolone Antibiotics

Subject Areas : Journal of Chemical Health Risks

Iman Fadhil Abdul-Husin 1 , Mohammed Sabri Abdul-Razzaq 2

1 - Department of Genetic Engineering, Faculty of Biotechnology, Al-Qasim Green University, Babylon, Iraq
2 - Department of Medical Microbiology, Faculty of Medicine, Babylon University, Babylon, Iraq

Received: 2021-07-01 Accepted : 2021-10-20 Published : 2021-12-01

Keywords: Antibiotic resistance, E.coli, Quinolone, Bacterial topoisomerases,

Abstract :

Multiple mutations in the quinolone-determining resistant regions of topoisomerase enzymes are usually associated with high levels of fluoroquinolone resistance in E. coli. Gyr A gene encodes (gyrase II) enzyme that changes topology by introducing transient double-stranded breaks into DNA and passing a second double-stranded DNA segment through the break before resealing it. The current investigation aims to reveal the correlation between functional gyrase enzyme and non-functional gyrase with level MIC of quinolone resistance under study. Patients with various illnesses such as urinary tract infections (UTIs), gastro enteritis (diarrhea) and wound infections were involved in the investigation. A total of 200 clinical samples were obtained. There are 30 samples diagnostic as E. coli clinical strain after identification by biochemical test, VITEK-2 compact system, and by molecular method using 16SrDNA marker. Antimicrobial susceptibility testing and minimal inhibition concentration (MIC) for Nalidixic acid, Norfloxacin, ciprofloxacin, levofloxacin, and Gatifloxacin was performed by broth microdilution method. According to manufacturing company (Topogen-USA); cell lysate are produced;then Bacterial gyrase assay (Topogen-USA) has been used to show the activity of gyrase enzyme by observation of negative supercoiling of E. coli DNA. Gyr A gene is detectable in all tested E. coli (100%); but There are only 16/30 isolates have active topoisomerase type2 with percentage 53.33% only; while; (14) isolates with non-functional gyrase enzyme since they not produce negative supercoiling DNA bands after gel electrophoreses with ethidium bromide stain. non-functional gyrase enzyme are associated with high E. coli resistance level against quinolone antimicrobial under study. MIC value for two groups are show significantly different concerning Nalidixic acid, Norfloxacin, while no significant difference concerning Levofloxacin and Gatifloxacin antibiotics.As a result, this research representsto be an effective method for determining the primary target of a quinolone drug in E. coli clinical isolates, and for the purification and characterization of DNA Gyrase so. E. coli are highly isolated from vagina samples 40% and then from urine samples (32%) followed by (24%) from wounds infections and (21%) from stool .There are correlation between the reduced sensitivity of quinolone under study and bacterial topoisomerases activity in E. coli isolates.

References:


  1. Bush N., Evans-Roberts K., Maxwell A., DNA topoisomerases. Eco Sal Plus doi: 10.1128/ecosalplus, ESP-0010-2014, 2015.
    2.
  2. Van den Broeck A., Carpini J., Leroy H., Diefendorff J.M., 2017. 19 How Much Effort Will I Put into My Work? It Depends on Your Type of Motivation. An introduction to work and organizational psychology: An international perspective, 354.
    3.
  3. Tchesnokova V., Radey M., Chattopadhyay S., Larson L., Weaver J. L., Kisiela D., Sokurenko E.V., 2019. Pandemic fluoroquinolone resistant Escherichia coli clone ST1193 emerged via simultaneous homologous recombinations in 11 gene loci. Proceedings of the National Academy of Sciences. 116(29), 14740-14748.
    4.
  4. Johnson J.R., Johnston B., Kuskowski M.A., Sokurenko E.V., Tchesnokova V., 2015. Intensity and Mechanisms of Fluoroquinolone Resistance within the H 30 and H 30Rx Subclones of Escherichia coli Sequence Type 131 Compared with Other Fluoroquinolone-Resistant E. coli. Antimicrobial Agents and Chemotherapy 59(8), 4471-4480.
    5.
  5. Shams E., Firoozeh F., Moniri R., Zibaei M., 2015. Prevalence of plasmid-mediated quinolone resistance genes among extended-spectrum β-Lactamase-producing Klebsiella pneumoniae human isolates in Iran. Journal of Pathogens. 2015, 1-7
    6.
  6. TopoGEN, DNA Gyrase Assay  Kit  Description  (TG1003). Florida.
    7.
  7. Forbes E.E., Shaw D.S., Dahl R.E., 2007. Alterations in reward-related decision making in boys with recent and future depression. Biological Psychiatry. 61(5), 633-639.
    8.
  8. Andrews J., Bradley J., Wise R., 1993. Comparison of ‘E’test with conventional agar MIC. Journal of Antimicrobial Chemotherapy. 31(5), 802-803.
    9.
  9. Institute C.a.L.S., 2020. Performance standard for antimicrobial susceptibility testing. 27th ed. CLSI supplement M100 Clinical and Laboratory Standards Institute. 34(5) 1-7.
    10.
  10. Ramírez Castillo F.Y.M.F.A.C., Avelar González F.J., Márquez Díaz F., Harel J., Guerrero Barrera A.L, 2018. An evaluation of multidrug resistant Escherichia coli isolates in urinary tract infectionsfrom Aguascalientes. Mexico: cross sectionalstudy.Ann ClinMicrobiol Antimicrob. 17 (34), 2-13.
    11.
  11. Varughese L.R., Rajpoot M., Goyal S., Mehra R., Chhokar V., Beniwal V., 2018. Analytical profiling of mutations in quinolone resistance determining region of gyrA gene among UPEC. PloS One. 13(1), e0190729.
    12.
  12. Lodish H., Berk A., Kaiser C.A., Kaiser C., Krieger M., Scott M.P., Bretscher A., Ploegh H., Matsudaira P., 2008. Molecular cell biology. Macmillan.
    13.
  13. Delmani F., Jaran A.S., TaraziY.A., Masaadeh H., Zaki O., Irbid T., 2017. Characterization of ampicillin resistant gene (blaTEM-1) isolated from E. coli in Northern Jordan. Asian J Biomed Pharm Sci. 7, 11-5.‏
    14.
  14. Saenz M., Hajer K, Mohamed S., Sana F., 2015. Occurrence of virulence genes in ESBL-producing Escherichia coli isolates from urine. Cell Infect Microbiol. 5(38), 322-344.
    15.
  15. 15. Orenstein R., Wong E.S., 1999. Urinary tract infections in adults. American Family physician. 59(5), 1225.‏
    16.
  16. Parvin K., Rahman N., Rahman M. M., Islam M.A., 2013. Isolation and Characterization of Microflora from Human Urine. Microbes and Health,. 2(1), 15-18.
    17.
  17. Otaiwi M.S., Tarrad J.K., AL-Yasari H.F., 2019. Prevalence of E. Coliserotypes among diarrheal patients in Hilla City, Iraq. Research Journal of Pharmacy and Technology. 12(7), 3347-3349.
    18.
  18. Jaloob Aljanaby A.A., Hashim Alfaham Q.M. 2017. Phenotypic and Molecular Characterization of some Virulence Factors in Multidrug Resistance Escherichia coli Isolated from Different Clinical Infections in Iraq. American Journal of Biochemistry and Molecular Biology. 7, 65-78.
    19.
  19. Hassan R., Tantawy M., Gouda N.A., Elzayat M.G., Gabra S., Nabih A., Diab A.A., El-Hadidi M., Bakry U., Shoeb M.R., 2020. Genotypic characterization of multiple drug resistant Escherichia coli isolates from a pediatric cancer hospital in Egypt. Scientific Reports. 10(1), 1-10.
    20.
  20. Al-Mayahie S.M., 2013. Phenotypic and genotypic comparison of ESBL production by vaginal Escherichia coli isolates from pregnant and non-pregnant women. Annals of Clinical Microbiology and Antimicrobials. 12(1), 1-7.
    21.
  21. Ali F.A., 2018. Distribution of CTX-M gene among Escherichia coli strains isolated from different clinical samples in Erbil City. Iraqi Journal of Biotechnology. 17(1), 1-7.
    22.
  22. Abdulqader H.H., Saadi A.T., 2019. The distribution of pathogens, risk factors and their antimicrobial susceptibility patterns among post-surgical site infection in Rizgari Teaching Hospital in Erbil/Kurdistan Region/Iraq. Journal of Duhok University. 22(1), 1-10.
    23.
  23. Adhikari K., Basnyat S., Shrestha B., 2020. Prevalence of Multidrug-Resistant and Extended-spectrum Betalactamase Producing Bacterial Isolates from Infected Wounds of patients in Kathmandu Model Hospital. Nepal Journal of Science and Technology. 19(1), 171-179.
    24.
  24. Opekun A.R., Gonzales S.A., Al-Saadi M.A., Graham D.Y., 2018. Brief report: Lactobacillus bulgaricus GLB 44 (Proviotic™) plus esomeprazole for Helicobacter pylori eradication: A pilot study. Helicobacter. 23(2), e12476.‏
    25.
  25. Pirko E.Y., Ali M.R., MR N., 2017. The Relationship between Phylogenic Typing and Antimicrobial Susceptibility Patterns forEscherichia coliIsolatedfrom UTIs atMany Hospitals in Baghdad City. Nursing National Iraqi Specialty. 30(2), 1-2.
    26.
  26. Ali I., Rafaque Z., Ahmed S., Malik S., Dasti J. I., 2016. Prevalence of multi-drug resistant uropathogenic Escherichia coli in Potohar region of Pakistan. Asian Pacific Journal of Tropical Biomedicine. 6(1), 60-66.
    27.
  27. Ekwealor P.A., Ugwu M.C., Ezeobi I., Amalukwe G., Ugwu B.C., Okezie U., Stanley C., Esimone C., 2016. Antimicrobial evaluation of bacterial isolates from urine specimen of patients with complaints of urinary tract infections in Awka, Nigeria. International Journal of Microbiology.2016,
    28.
  28. Zahedi Z., Hakemi Vala M., Bagheri Bejestani F., 2018. Contribution of GyrA and QnrA genes in ciprofloxacin resistant Ecsherichia coli isolates from patients with urinary tract infections of Imam Khomeini Hospital of Tehran. Archives of Pediatric Infectious Diseases. 6(4), 1-7.
    29.
  29. Stracy M., Wollman A.J.M., Kaja E., Gapinski J., Lee J.-E., Leek V.A., McKie S.J., Mitchenall L.A., Maxwell A., Sherratt D.J., 2019. Single-molecule imaging of DNA gyrase activity in living Escherichia coli. Nucleic Acids Research. 47(1), 210-220.
    30.
  30. Marcusson L.L., Frimodt-Møller N., Hughes D., 2009. Interplay in the selection of fluoroquinolone resistance and bacterial fitness. PLoS Pathogens. 5(8), e1000541.
    31.
  31. Majlesi A., Kakhki R. K., Nejad A. S. M., Mashouf R.Y., Roointan A., Abazari M., Alikhani M.Y., 2018. Detection of plasmid-mediated quinolone resistance in clinical isolates of Enterobacteriaceae strains in Hamadan, West of Iran. Saudi Journal of Biological Sciences. 25(3), 426-430.
    32.
  32. da Silva G.J., Mendonça N., 2012. Association between antimicrobial resistance and virulence in Escherichia coli. Virulence. 3(1), 18-28.
    33.
  33. Webber M.A., Ricci V., Whitehead R., Patel M., Fookes M., Ivens A., Piddock L.J., 2013. Clinically relevant mutant DNA gyrase alters supercoiling, changes the transcriptome, and confers multidrug resistance. MBio. 4(4), e00273-00213.
    34.
  34. Gomig F., Galvão C.W., Freitas D.L.d., Labas L., Etto R.M., Esmerino L.A., Lima M.A.d., Appel M.H., Zanata S.M., Steffens M.B.R., 2015. Quinolone resistance and ornithine decarboxylation activity in lactose-negative Escherichia coli. Brazilian Journal of Microbiology. 46, 753-757.
    35.
  35. Gruger T., Nitiss J. L., Maxwell A., Zechiedrich E. L., Heisig P., Seeber S., Pommier Y., Strumberg D., 2004. A mutation in Escherichia coli DNA gyrase conferring quinolone resistance results in sensitivity to drugs targeting eukaryotic topoisomerase II. Antimicrobial Agents and Chemotherapy. 48(12), 4495-4504.
    36.
  36. Lorestani R.C., Akya A., Elahi A., 2018. The mutations of topoisomerase genes and their effect on resistance to fluoroquinolones in extended-spectrum β-lactamase-producing Escherichia coli. Jundishapur Journal of Natural Pharmaceutical Products. 13(1)1-6
    37.
  37. Ogbolu D.O., Daini O., Ogunledun A., Alli A.T., Olusoga-Ogbolu F., Webber M., 2012. Effects of gyrA and parC mutations in quinolones resistant clinical gram negative Bacteria from Nigeria. African Journal of Biomedical Research. 15(2), 97-104.
    38.
  38. Redgrave L.S., Sutton S.B., Webber M.A., Piddock L.J., 2014. Fluoroquinolone resistance: mechanisms, impact on bacteria, and role in evolutionary success. Trends in Microbiology., 22(8), 438-445.
    39.
  39. Shenagari M., Bakhtiari M., Mojtahedi A., Roushan Z.A., 2018. High frequency of mutations in gyrA gene associated with quinolones resistance in uropathogenic Escherichia coli isolates from the north of Iran. Iranian Journal of Basic Medical Sciences. 21(12), 1226.
    40.
  40. Haggag Y., Saafan A., El-Gendy A., Hefzy E., AbdelGhani S., 2020. Molecular Characterization of Quinolone Resistant Urinary Isolates of Escherichia coli. J Pure Appl Microbiol. 14(2), 1269-1277.
    41.
  41. Kim H.B., Wang M., Park C.H., Kim E.C., Jacoby G.A., Hooper D.C., 2009. oqxAB encoding a multidrug efflux pump in human clinical isolates of Enterobacteriaceae. Antimicrobial Agents and Chemotherapy. 53(8), 3582-3584.
    42.
  42. Cavaco L., Aarestrup F.M., 2009. Evaluation of quinolones for use in detection of determinants of acquired quinolone resistance, including the new transmissible resistance mechanisms qnrA, qnrB, qnrS, and aac (6′) Ib-cr, in Escherichia coli and Salmonella enterica and determinations of wild-type distributions. Journal of Clinical Microbiology. 47(9), 2751-2758.
    43.
  43. Oviaño M., Rodríguez-Martínez M.J., PascualA.andBou G., 2017. Rapid detection of the plasmid-mediated quinolone resistance determinant AAC(6′)-Ib-cr in Enterobacteriaceae by MALDI-TOFMS analysis. Journal of Antimicrobial Chemotherapy. 72(4), 1074–1080.
    44.
  44. Hopkins K.L., Davies R.H., Threlfall E.J., 2005. Mechanisms of quinolone resistance in Escherichia coli and Salmonella: recent developments. International Journal of Antimicrobial Agents. 25(5), 358-373.
    45.
  45. Abbasi H., Ranjbar R., 2018. The prevalence of quinolone resistance genes of A, B, S in Escherichia coli strains isolated from three major hospitals in Tehran, Iran. Central European Journal of Urology. 71(1), 129.
    46.
  46. Pfeiffer E.S., Hiasa H., 2007. Determination of the primary target of a quinolone drug and the effect of quinolone resistance-conferring mutations by measuring quinolone sensitivity based on its mode of action. Antimicrobial Agents and Chemotherapy. 51(9), 3410-3412.
    47.
  47. Cesaro A., Bettoni R.R.D., Lascols C., Mérens A., Soussy C.J., Cambau E., 2008. Low selection of topoisomerase mutants from strains of Escherichia coli harbouring plasmid-borne qnr genes. Journal of Antimicrobial Chemotherapy. 61(5), 1007-1015.
    48.
  48. Yousefi S., Mojtahedi A., Shenagari M., 2018. A Survey of gyrA Target-Site Mutation and qnr Genes among Clinical Isolates of Escherichia coli in the North of Iran. Jundishapur Journal of Microbiology. 11(9).
    49.
  49. Drago L., Nicola L., Mattina R., De Vecchi E., 2010. In vitro selection of resistance in Escherichia coli and Klebsiella spp. at in vivo fluoroquinolone concentrations. BMC Microbiology. 10(1), 1-7.
    50.
  50. Minarini L.A., Darini A.L.C., 2012. Mutations in the quinolone resistance-determining regions of gyrA and parC in Enterobacteriaceae isolates from Brazil. Brazilian Journal of Microbiology. 43(4), 1309-1314.
    51.
  51. Bhatnagar K., Wong A., 2019. The mutational landscape of quinolone resistance in Escherichia coli. PloS One. 14(11), e0224650.
    52.
  52. Lu T., Zhao X., Drlica K., 1999. Gatifloxacin activity against quinolone-resistant gyrase: allele-specific enhancement of bacteriostatic and bactericidal activities by the C-8-methoxy group. Antimicrobial Agents and Chemotherapy. 43(12), 2969-2974.
    53.

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