Molecular characterization of β-lactam resistance genes in clinical bacterial isolates of the Enterobacteriaceae family
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Published:
Apr 23, 2021
Keywords:
allelic variant, bacterial resistance, Enterobacteriaceae, extended spectrum betalactamases (ESBL), resistance genes
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Abstract
Infections developed by extended spectrum β-lactamase-producing Enterobacteriaceae (ESBL) are associated with high mortality and morbidity rates in hospital settings due to their ability to hydrolyze β-lactam antibiotics. The objective of this study was to characterize the genes that confer resistance to β-lactams in Enterobacteriaceae obtained from a tertiary hospital in the city of Quito. 153 Enterobacteriaceae were collected and the species was identified with biochemical tests. The study selected the isolates that presented ESBL enzyme production analyzed by the double disk synergy method and the VITEK 2 automated system provided by the hospital. Of the 22 selected isolates, 19 were identified as Escherichia coli and 3 as Klebsiella oxytoca. The ability of the isolates to produce carbapanemase enzymes was determined with the Triton Hodge Test (THT), showing that no isolate had this capacity. The identification of resistance genes used a polymerase chain reaction and used specific primers for each gene encoding ESBL (blaCTX-M, blaTEM, blaSHV) and carbapenemase enzymes (blaKPC, blaIMP, blaVIM, blaNDM). The identification of the allelic variant reported that 11/22 isolates presented the blaCTX-M-15 gene and 4/22 the blaTEM-1 gene. None of the isolates presented genes for resistance to carbapenems.
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Coral D, Yauri Bucheli MF, Alcocer I. Molecular characterization of β-lactam resistance genes in clinical bacterial isolates of the Enterobacteriaceae family. REMCB [Internet]. 2021Apr.23 [cited 2024Jul.3];42(1). Available from: https://remcb-puce.edu.ec/remcb/article/view/886
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References
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Elena A, Cejas D, Magariños F, Jewtuchowicz V, Facente A, Gutkind G, Di Conza J, Radice M. 2018. Spread of Clonally RelatedEscherichia coli Strains Harboringan IncA/C1 Plasmid Encoding IMP-8 and Its Recruitment intoan Unrelated MCR-1-Containing Isolate. Antimicrobial agents and chemotherapy. 62(6): 02414-02417.
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Guillén, R. et al. 2016. Detección molecular de belactamasas de espectro extendido (BLEE) en enterobacterias aisladas en Asunción. Mem Inst Investig Cienc Salud. 14(1):8–16.
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O’Hara J, Hu F, Ahn C, Nelson J, Rivera J, Pasculle A, Doi Y. 2014. Molecular epidemiology of KPC-producing Escherichia coli: Occurrence of ST131-fimH30 subclone harboring pKpQIL-like IncFIIk plasmid. Antimicrob Agents Chemother. 58(7):4234–4237.
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Pasteran F, Gonzalez L, Albornoz E, Bahr G, Vila A, Corso A. 2016. Triton Hodge test: improved protocol for modified Hodge test for enhanced detection of NDM and other carbapenemase producers. J Clin Microbiol 54:640 –649. doi:10.1128/JCM.01298-15.
Poirel L, Naas T, Nicolas D, Collet L, Bellais S, Cavallo J, Nordmann P. 2000. Characterization of VIM-2, a Carbapenem-Hydrolyzing Metallo-β-Lactamase and Its Plasmid and Integron Borne Gene from a Pseudomonas aeruginosa Clinical Isolate in France. Antimicrobial agents and chemotherapy. 44: 891-897.
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BD Diagnostics. 2009. Difco and BBL Manual Manual of Microbiological Culture Media 2nd Edition.
Bauer A, Kirby W, Sherris J, TurckM. 1966. Antibiotic Susceptibility Testing By A Standardized Single Disk Method. Am J Clin Pathol. 45(4):493–496.
Bevan E, Jones A, Hawkey P. 2017. Global epidemiology of CTX-M β-lactamases: Temporal and geographical shifts in genotype. J Antimicrob Chemother. 72(8):2145–2155.
Blanco V, Maya J, Correa A, Perenguez M, Muñoz J, Motoa G, Pallares C, Rosso F, Matta L, Celis Y, et al. 2016. Prevalencia y factores de riesgo para infecciones del tracto urinario de inicio en la comunidad causadas por Escherichia coli productor de betalactamasas de espectro extendido en Colombia. Enferm Infecc Microbiol Clin. 34(9):559–565.
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Cantón R, González-Alba J, Galán J. 2012. CTX-M enzymes: Origin and diffusion. Front Microbiol. 3(APR).
(CDC) Centers for Disease Control and Prevention. 2013. Vital Signs : Carbapenem-Resistant Enterobacteriaceae. MMWR Morb Mortal Wkly Rep. 62(9):165–170.
Chen L, Freeman J, Nicholson B, Keiger A, Lancaster S, Joyce M, Woods C, Cook E, Adcock L, Louis S, et al. 2014. Widespread dissemination of CTX-M-15 genotype extended-spectrum-β- lactamase-producing enterobacteriaceae among patients presenting to community hospitals in the southeastern United States. Antimicrob Agents Chemother. 58(2):1200–1202.
Cho H, Uehara T, Bernhardt T. 2014. Beta-lactam antibiotics induce a lethal malfunctioning of the bacterial cell wall synthesis machinery. Cell. 159(6):1300–1311.
(CLSI) Clinical and Laboratory Standards Institute. 2018. Clinical and Laboratory Standards Institute antimicrobial susceptibility testing standards M02, M07, and M11.
Codjoe F, Donkor E. 2017. Carbapenem Resistance: A Review. Med Sci. 6(1):1.
Colquechagua F, Sevilla C, Gonzales E. 2015. Artículo Original de espectro extendido en muestras fecales en el enterobacteriaceae in fecal samples at the national. Rev Peru Med Exp Salud Publica. 32(1):26–32.
D’Andrea M, Arena F, Pallecchi L, Rossolini G. 2013. CTX-M-type β-lactamases: A successful story of antibiotic resistance. Int J Med Microbiol. 303(6–7):305–317.
Calva D, Toledo Z, Ochoa S, Arévalo A, Ausili A. 2016. Detection and molecular characterization of β-lactamase genes in clinical isolates of Gram-negative bacteria in Southern Ecuador. Brazilian J Infect Dis. 20(6):627–630.
Díaz V. 2017. Caracterización molecular de genes de resistencia a β-lactámicos en aislados clínicos de Escherichia coli provenientes de urocultivos y pruebas de inhibición con péptidos de Boana rosenbergi y Rana sp. [tesis]. [Quito (UIO)]: Pontificia Universidad Católica del Ecuador.
Ding H, Liu B, Gao Y, Zhong X, Duan S, Yuan L. 2018. Divergence of affinities, serotypes and virulence factor between CTX-M Escherichia coli and non-CTX-M producers. Poult Sci. 97(3):980–985.
Doi Y, Iovleva A, Bonomo RA. 2017. The ecology of extended-spectrum β-lactamases (ESBLs) in the developed world. J Travel Med. 24(1):S44–S51.
Dortet L, Nordmann P, Poirel L. 2012. Association of the Emerging Carbapenemase NDM-1 with aBleomycin Resistance Protein inEnterobacteriaceaeandAcinetobacter baumannii. Antimicrobial agents and chemotherapy. 56(4):1693.
Elena A, Cejas D, Magariños F, Jewtuchowicz V, Facente A, Gutkind G, Di Conza J, Radice M. 2018. Spread of Clonally RelatedEscherichia coli Strains Harboringan IncA/C1 Plasmid Encoding IMP-8 and Its Recruitment intoan Unrelated MCR-1-Containing Isolate. Antimicrobial agents and chemotherapy. 62(6): 02414-02417.
Escandón K, Reyes S, Gutiérrez S, Villegas M. 2017. The epidemiology of carbapenemases in Latin America and the Caribbean. Expert Rev Anti Infect Ther. 15(3):277–297.
Fadil Saedii A, Abdelraheim A, Abdel Aziz A, Swelam S. 2017. ESBL-Producing E.Coli and Klebsiella among Patients Treated at Minia University Hospitals. J Anc Dis Prev Remedies. 05(02).
Fatemeh F, Hamed A, Ali H, Hossein G, Latif G, Parastoo H. 2016. Antibiotic susceptibility patterns in CTX-M-15-producing Enterobacteraceae isolated from healthy Afghan refugees in Iran. African J Microbiol Res. 10(11):357–362.
Flores A, Walker J, Caparon M, Hultgren SJ. 2015. Urinary tract infections: Epidemiology, mechanisms of infection and treatment options. Nat Rev Microbiol. 13(5):269–284.
Guillén, R. et al. 2016. Detección molecular de belactamasas de espectro extendido (BLEE) en enterobacterias aisladas en Asunción. Mem Inst Investig Cienc Salud. 14(1):8–16.
Guzmán-Blanco M, Labarca J, Villegas M, Gotuzzo E. 2014. Extended spectrum β-lactamase producers among nosocomial Enterobacteriaceae in Latin America. Brazilian J Infect Dis. 18(4):421–433.
INEC (2017). Programa Nacional de Estadística 2017-2021. Instituto Nacional de Estadística y Censos, Quito-Ecuador.
Iñiguez D, Zurita J, Alcocer I, Ortega D, Gómez AM, Maldonado L. 2012. Klebsiella pneumoniae productora de carbapenemasa tipo KPC-2: primer reporte en el Ecuador. Rev la Fac Ciencias Médicas. 37(1–2):40–43.
Kaluzna E, Zalas-Wiecek P, Gospodarek E. 2014. Comparison of detection methods for extended-spectrum beta-lactamases in Escherichia coli strains. Postepy Hig Med Dosw. 68(June):808–813.
Kang H, Wang L, Li Y, Lu Y, Fan W, Bi R, Qian H, Gu B. 2019. Dissemination of Multidrug-Resistant Shigella flexneri and Shigella sonnei with Class 1, Class 2, and Atypical Class 1 Integrons in China. Microb Drug Resist. 00(00):mdr.2018.0229.
Kelly A, Mathema B, Larson E. 2017. Carbapenem-resistant Enterobacteriaceae in the community: a scoping review. Int J Antimicrob Agents. 50(2):127–134.
López L. 2014. Papel del ambiente hospitalario y los equipamientos en la transmisión de las infecciones nosocomiales. Enferm Infecc Microbiol Clin. 32(7):459–464.
López D, Torres M, Prada C. 2016. Genes de resistencia en bacilos Gram negativos: Impacto en la salud pública en Colombia. Univ y Salud. 18(1):190.
Macy E, Contreras R. 2015. Adverse reactions associated with oral and parenteral use of cephalosporins: A retrospective population-based analysis. J Allergy Clin Immunol. 135(3):745-752.e5.
Mathers A, Peirano G, Pitout J. 2015. The role of epidemic resistance plasmids and international high- risk clones in the spread of multidrug-resistant Enterobacteriaceae. Clin Microbiol Rev. 28(3):565–591.
Morones I, Salgado T, Gonzaga T, Matamoros A, Terán J, Arteaga S, Castro L, Reyes A, López D, Meza D. 2016. Enterobacterias con betalactamasas de espectro extendido en hemocultivos y urocultivos. Med Interna Mex. 32(4):381–387.
Numanovic F, Hukic M, Delibegovic Z, Tihic N, Pasic S. 2013. Comparison of double disk synergy test , VITEK 2 and Check- MDR CT102 for detection of ESBL producing isolates. Acta Medica Acad 2013. 42(1):15–24.
O’Hara J, Hu F, Ahn C, Nelson J, Rivera J, Pasculle A, Doi Y. 2014. Molecular epidemiology of KPC-producing Escherichia coli: Occurrence of ST131-fimH30 subclone harboring pKpQIL-like IncFIIk plasmid. Antimicrob Agents Chemother. 58(7):4234–4237.
Oliveira M, Oliveira C, Gonçalves K, Santos M, Tardelli A, Nobre V. 2015. Enterobacteriaceae resistant to third generation cephalosporins upon hospital admission: Risk factors and clinical outcomes. Brazilian J Infect Dis. 19(3):239–245.
Organización Mundial de la Salud (OMS) [Internet]. 2018. Bangkok. Datos recientes revelan los altos niveles de resistencia a los antibióticos en todo el mundo; [updated 2018 Ene 30; cited 2020 Nov 20]. Disponible en: https://www.who.int/es/news/item/29-01-2018-high-levels-of-antibiotic-resistance-found-worldwide-new-data-shows
Pachay S. 2018. Las infecciones bacterianas y su resistencia a los antibióticos. Caso de estudio: Hospital Oncológico “Dr. Julio Villacreses Colmont Solca”, Portoviejo. Univ y Soc. 10(3):134–141.
Pasteran F, Gonzalez L, Albornoz E, Bahr G, Vila A, Corso A. 2016. Triton Hodge test: improved protocol for modified Hodge test for enhanced detection of NDM and other carbapenemase producers. J Clin Microbiol 54:640 –649. doi:10.1128/JCM.01298-15.
Poirel L, Naas T, Nicolas D, Collet L, Bellais S, Cavallo J, Nordmann P. 2000. Characterization of VIM-2, a Carbapenem-Hydrolyzing Metallo-β-Lactamase and Its Plasmid and Integron Borne Gene from a Pseudomonas aeruginosa Clinical Isolate in France. Antimicrobial agents and chemotherapy. 44: 891-897.
Promega C. 2010. Isolation of Genomic DNA from Whole Blood Wizard ® Genomic DNA Purification Kit. :1123–1126.
Ranjbar R, Ardashiri M, Samadi S, Afshar D. 2018. Distribution of extended-spectrum β-lactamases (ESBLs) among salmonella serogroups isolated from pediatric patients. Iran J Microbiol. 10(5):294–299.
Rhodes N, Richardson C, Heraty R, Liu J, Malczynski M, Qi C, Scheetz M. 2014. Unacceptably high error rates in Vitek 2 testing of cefepime susceptibility in extended-spectrum-β-lactamase-producing Escherichia coli. Antimicrob Agents Chemother. 58(7):3757–3761.
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