Klinik E.coli İzolatlarında Virulans Faktor Genlerinin ve Geniş Spekturumlu Beta Laktamazların Araştırılması

Year 2020, Volume: 10 Issue: 1, 122 - 127, 15.01.2020

Azer Özad Düzgün Esma Akyıldız Ayşegül Saral Tuba Köse

https://doi.org/10.17714/gumusfenbil.571958

Cited By: 1

Abstract

Bu çalışmanın amacı, 83 klinik E.coli izolatında pap, hly,
cnf, sfa,aer ve afa virulans
genlerinin ve geniş spekturumlu beta laktamaz direnç genlerinin varlığını
araştırmaktır.  83 E. coli izolatında sfa, pap,
hly, cnf, aerve afa virülans
genleri ve TEM, SHV, CTX-M1 ve CTX-M9 geniş spekturumlu beta laktamaz genleri polimer
zincir reaksiyonu (PZR) metodu ile araştırılmıştır. 83 izolatın, sefiksim,
trimethoprim/sulfametaksazol, piperasilin/tazobaktam, amoksisilin/klavulanik
asit, ampisilin, seftriakson, sefuroksim, sefuroksim aksetil, siprofloksasin,
fosfomisin, gentamisin ve seftazidime karşı direnç oranları sırasıyla;%65, %54,
%19, %38, %74, %75, %72, %75, %68, %2, %30, %49 olarak belirlenmiştir. CTX-M1,
araştırılan genişlemiş spektrumlu beta-laktamaz (GSBL) genleri arasında 83
suşun 51’inde belirlenmesiyle en sık görülen GSBL olmuştur. TEM tipi beta
laktamaz 33 suşta, CTX-M9 ve SHV 1 izolatta tespit edilmiştir. İzolatlarda en
yaygın virülans geni, 40 suşta tespit edilmesiyle aer geni, bunu izolatların 15'inde tespit edilen cnf geni takip etmiştir. Sfa, pap, afavehlyvirulans genleri sırasıyla izolatların %15.6'sında, %15.6'sında,
% 9.6'sında ve %12.04’ünde görülmüştür. Ayrıca 83 E.coliizolatında on farklı virulans faktör gen paterni tespit
edilmiştir. Virulans gen kombinasyonları, 27 suştaaer, 6 suştacnf-sfa-pap, 1
suştaaer-pap, 2 suştaaer-afa, 6 suştaafa,
3 suştasfa, 9 suştaaer-cnf-hly, 4 suştasfa-pap, 1 suştahly-papve
1 suştapap olarak gözlemlenmiştir. Sonuç
olarak, İYE ile ilişkili bakteriyel patojenitenin araştırılması daha iyi bir
tıbbi müdahaleye katkı sağlayacaktır. 

Keywords

Direnç, E.coli, Virulans

Supporting Institution

GÜBAP

Project Number

18.119.03.02

Thanks

Bu çalışma 18.119.03.02 nolu Gümüşhane Üniversitesi, Bilimsel Araştırma Birimi ile desteklenmiştir.

Investigation of Virulence Factor Genes and Extended Spekturum Beta Lactamases in Clinical E.coli Isolates

Abstract

The aim of this study
was to investigate the presence of pap, hly, cnf, sfa, aer and afa virulans genes
and extended-spectrum beta lactamase resistance genes in 83 clinical E.coli
isolates. In 83 E. coli isolates, sfa, pap, hly, cnf, aer and afa virulence
genes and TEM-SHV, CTX-M1 and CTX-M9 wide spectrum beta-lactamase genes were
investigated by polymerase chain reaction (PCR) method. Resistance rates of 83
isolates to cefixim, trimethoprim/sulfamethoxazole, piperacillin/tazobactam,
amoxicillin/clavulanic acid, ampicillin, ceftriaxone, cefuroxime, cefuroxime
axetil, ciprofloxacin, fosfomycin, gentamicin and ceftazidime antibiotics were
65%, 54%, 19%,38, 74%, 75%, 72%, 75%, 68%, 2%, 30%, 49%, respectively. CTX-M1
was the most common extended spectrum beta-lactamase (ESBL) among the ESBL
genes investigated in 51 of 83 strains. TEM type beta-lactamase was detected in
33 strains, CTX-M9 and SHV 1 isolates. The most common virulence gene was aer
gene and detected in 40 strains, followed by the cnf gene detected in 15 of the
isolates. The pap, hly, afa and sfa genes were determined in 15.6%, 12.04%,
9.6% and 15.6% of the isolates, respectively. In addition, 10 different
virulence factor gene patterns were observed in 83 E.coli isolates. Virulence
gene combinations were observed in 27 strains of aer, 6 strains of cnf-sfa-pap,
1 strains of aer-pap, 2 strains of aer-afa, 6 strains of afa, 3 strain of sfa,
9 strain of aer-cnf-hly, 4 isalates of sfa-pap, 1 strain of hly-pap and 1
strains of pap genes. In conclusion, investigating bacterial pathogenicity
associated with UTI will contribute to better medical intervention.

Keywords

Resistance, E.coli, Virulence

Project Number

18.119.03.02

References

• Abe, C.M., Salvador, F.A., Falsetti, I.N., Vieira, M.A, Blanco, J., Blanco, J.E., Blanco, M., Machado, A.M., Elias, W.P., Hernandes, R.T., Gomes, T.A.,2008.Uropathogenic Escherichia coli (UPEC) strains may carry virulence properties of diarrhoeagenic E. coli. FEMS Immunol Med Microbiol, 52,397–406.
• Alyamani, E.J., Khiyami, M.A., Booq, R.Y., Alnafjan, B.M., Altammami, M.A., Bahwerth,FS.,2015.Molecular characterization of extended-spectrum beta-lactamases (ESBLs) produced by clinical isolates of Acinetobacterbaumannii in Saudi Arabia. Ann ClinMicrobiolAntimicrob,20,38.
• Arabi, S., Tohidi, F., Naderi, S., Nazemi, A., Jafarpour, M., Naghshbandi, R.,2012. The common fimbarie genotyping in Uropathogenic Escherichia coli. Ann Biol Res, 3,4951–4954.
• Bektaş, A., Güdücüoğlu, H., Gürsoy, N.C., Berktaş, M., Gültepe, B.S., Parlak, M., Otlu, B. ve Mehmet Sait Tekerekoğlu, M.S., 2018. Genişlemiş spektrumlu beta-laktamaz (GSBL) üreten Escherichia coli ve Klebsiella pneumoniae suşlarının gsbl genlerinin araştırılması, Flora İnfeksiyon Hastalıkları ve Klinik Mikrobiyoloji Dergisi, 23,116-123.
• Bien, J., Sokolova, O., Bozko, P., 2012. Role of uropathogenic Escherichia coli virulence factors in development of urinary tract infection and kidney damage. Int J Nephrol, 2012,1–15.
• Bush, K., Jacoby, G.A.,2010.Updated functional classification of β-lactamases.Antimicrob Agents Chemother,54,969–976.
• Cicek Copur, A., Saral, A., OzadDuzgun, A., Yasar, E., Cizmeci, Z., OzlemBalci, P., Sari, F., Firat, M., Altintop,Y.A., Ak, S., Caliskan, A., Yildiz, N., Sancaktar, M., EsraBudak, E., Erturk, A., BirolOzgumus, O., Sandalli, C., 2013.Nationwide study of Escherichia coli producing extended-spectrum β-lactamases TEM, SHV and CTX-M in Turkey.J Antibiot (Tokyo), 66,647-50.
• Denk, A. ve Sağmak Tartar, A., 2015. İdrar kültürlerinden izole edilen toplum kökenli Escherichia coli suşlarında antibiyotik direnci, Fırat Üniversitesi Sağlık Bilimleri Dergisi, 29, 51-55.
• Doğru, A., Üçışık, A.C., Sargın, F., Aydın, Ö., Ergen, P. ve Tükenmez Tigen, E., 2014. İdrar örneklerinde üretilen Escherichia coli suşlarında genişlemiş spektrumlu beta-laktamaz varlığı ve antibiyotik duyarlılıkları, Göztepe Tıp Dergisi, 29,219-224. Duman, Y., Güçlüer, N., Serindağ, A. ve Tekerekoğlu, M.S., 2010. Escherichia coli suşlarında antimikrobiyal duyarlılık ve genişlemiş spektrumlu-βeta laktamaz (GSBL) varlığı, Fırat Tıp Dergisi, 15, 197-200.
• Düzgün, Özad A., Okumuş, F., Saral, A., Çiçek, A.Ç., Cinemre, S., 2019. Determination of antibiotic resistance genes and virulence factors in Escherichia coli isolated from patients with urinary tract infection from Turkey. Revista da SociedadeBrasileira de Medicina Tropical.Basımda.
• Emody, L., Kerényi, M., Nagy, G., 2003. Virulence factors of uropathogenic Escherichia coli. International Journal of Antimicrobial Agents, 22,29-33.
• Eucast Version 9.0: The European Committee on Antimicrobial Susceptibility Testing. Breakpoint tables for interpretation of MICs and zone diameters. Version 9.0, 2019. http://www.eucast.org
• Firoozeh, F., Saffari, M., Neamati, F., Zibaei, M.,2014. Detection of virulence genes in Escherichia coli isolated from patients with cystitis and pyelonephritis. International Journal of Infectious Diseases 29, 219–222.
• Fluit, A.C., Visser, M.R, Schmitz, F.J., 2001.Molecular detection of antimicrobial resistance.ClinMicrob Rev 14,836–871
• Johnson, J.R.,1991. Virulence factors in Escherichia coli urinary tract infection. ClinMicrobiol Rev, 4,80- 128.
• Johnson, J.R., Kuskowski, M.A., Obryan, T., Colodner, R., Raz, R., 2006. Virulence genotype and phylogenetic origin in relation to antibiotic resistance profile among Escherichia coli urine sample isolates from Israeli women with acute uncomplicated cystitis. Antimicrob Agents Chemother, 49,26–31.
• Kaçmaz, B., Aksoy, A. ve Sultan, N., 2007. İdrar örneklerinden izole edilen Escherichia coli izolatlarında oral antibiyotiklere karşı direncin araştırılması, Türk Hijyen ve Deneysel Biyoloji Dergisi, 64, 11-15.
• Karimian, A., Momtaz, H., Mahbobe, M.M., 2012. Detection of uropathogenic Escherichia coli virulence factors in patients with urinary tract infections in Iran. Afr J Microbiol Res, 6,6811–6816.
• Kuzucu, Ç., Yetkin, F., Görgeç, S. ve Yasemin Ersoy, Y., 2011. Genişlemiş spektrumlu beta-laktamaz üreten Escherichia coli ve Klebsiella spp. suşlarının ertapenem ve diğer karbapenemlere karşı duyarlılıklarının araştırılması, Mikrobiyoloji Bülteni, 45, 28-35.
• Momtaz, H., Karimian, A., Madani, M., SafarpoorDehkordi, F., Ranjbar, R., Sarshar, M., Souod, N.,2013. Uropathogenic Escherichia coli in Iran: Serogroup distributions, virulence factors and antimicrobial resistance properties. Annals of Clinical Microbiology and Antimicrobials, 29,12-8.
• Oliveira, F.A., Paludo, K.S., Arend, L.N., Farah, S.M., Pedrosa, F.O., Souza, E.M., Surek, M., Picheth, G., Fadel-Picheth,C.M.,2011. Virulence characteristics and antimicrobial susceptibility of uropathogenic Escherichia coli strains. Genet Mol Res, 10,4114–4125.
• Paterson, D.L., Bonomo, R.A., 2005. Extended-spectrum β-lactamases: a clinical update. ClinMicrobiol Rev, 18,657–686.
• Polse, R.F., Yousif, S.Y. ve Assafi, M.S., 2016. Prevalence and molecular characterization of extended spectrum betaLactamases-producing uropathogenic Escherichia coli isolated in Zakho, Iraq, Journal of Microbiology and Infectious Diseases, 6, 163-167.
• Rahdar, M., Rashki, A., Miri, H.R., RashkiGhalehnoo, M.,2015. Detection of pap, sfa, afa, foc, and fimAdhesin-Encoding Operons in Uropathogenic Escherichia coli Isolates Collected From Patients With Urinary Tract Infection.Jundishapur J Microbiol, 8,e22647. Rawat, D., Nair, D., 2010. Extended-spectrum β-lactamases in Gram negative bacteria. J Glob Infect Dis, 2,263.
• Sana, T., Rami, K., Racha, B., Fouad, D., Marcel, A., Hassan, M., Sani, H. ve Monze, H., 2011. Detection of genes TEM, OXA, SHV and CTX-M in 73 clinical isolates of Escherichia coli producers of extended spectrum beta-lactamases and determination of their susceptibility to antibiotics, The Internatıonal Arabıc Journal, 1,1:5.
• Shakya, P., Shrestha, D., Maharjan, E., Sharma, V.K., Paudyal, R., 2017.ESBL Production AmongE. coli and Klebsiella spp. Causing Urinary Tract Infection: A Hospital Based Study. Open Microbiol J,28,23-30.
• Stordeur, P., Bree, A., Mainil, J., Moulin-Schouleur, M.,2004. Pathogenicity of pap-negative avian Escherichia coli isolated from septicaemic lesions. Microbes Infect,6,637–45.
• Subashchandrabose, S., Mobley, H.L.T., 2015.Virulence and Fitness Determinants of Uropathogenic Escherichia coli.MicrobiolSpectr,doi: 10.1128/microbiolspec.UTI-0015-2012.
• Sujatha, R., Kumar, A. ve Kumar, A., 2017. Molecular characterization of TEM, SHV and CTX-M extended spectrum beta lactamase among Escherichia coli and Klebsiella pneumoniae ın urinary ısolates, Journal of Dental and Medical Sciences, 16, 76-81.
• Tarchouna, M., Ferjani, A., Ben-Selma, W., Boukadida, J., 2013. Distribution of uropathogenic virulence genes in Escherichia coli isolated from patients with urinary tract infection. Int J Infect Dis, 17,e450-3.
• Terlizzi, M.E., Gribaudo, G., ve Maffei, M.E., 2017. UroPathogenic Escherichia coli (UPEC) infections: virulence factors, bladder responses, antibiotic, and non-antibiotic antimicrobial strategies, Of The Journal Frontiers in Microbiology, 8,1-23.
• Yun, K.W., Kim, H.Y., Park, H.K., Kim, W., Lim, I.S.,2014.Virulencefactors of uropathogenicEscherichiacoli of urinarytractinfectionsandasymptomaticbacteriuria in children.J MicrobiolImmunolInfect,47,455–61.