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مقاله پژوهشي
1116
Escherichiaيها زولهيادر CTX-M-1 ژن يفراوان
coli نمونه در ف يالط عيم بتاالکتاماز وسيمولد آنز
اي زنجيره واکنش روش با ينيبالهاي
در ياسوج PCRپليمراز
*1يد خسروانيد عبدالمجيس 3اييسيده مرضيه طباطب1يفيصغر شر1د سجادخرم روزيس 2ينيحس يمهر ،1شونديزهرا خو
ده يچک
هدف و
. (ESBLs) E. coli
CTX-M-1Escherichia coli
PCR
روش بررسي
CLSI
.CTX-M1ESBLPCR
. CTX-M-1 E.coli PCR ها: افتهي
PCR
CTX-M-1
.
E.coli يريگ جهينت
E. coli CTX-M-1کليدي: يها هواژ
،يد خسروانيد عبدالمجيس نويسنده مسئول:*Email: [email protected]
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مقدمه
.
E. coli
Escherichia coli
.
Beta-lactamase-enzymes
Extended
Spectrum Bete-Lactamase
ABCD
A
TEMSHV CTX-M
ESBL
SHV TEM
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ESBLs
CTX-M
)CTX-M-9 CTX-M-8 CTX-M-25CTX-M-2 (CTX-M-1
.E. coli
ESBL
E. coli
.
.
CTX-M
ESBL
.
E.coli
CTX-M-1PCR
يروش بررس
E. coli
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EMB
MRVPSIMTSI
Escherichia coli
E. coli
CLSI (Clinical & Laboratory Standards Institute) M100-
A20 (2010)
CTX-M-1
DNA boiling
PCR
SPSS
ها يافته
E. coli
.
.
Combined disk
E. coli
.
CTX-M-1
E. coliESBLs
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CTXM-1ژن يمرهايپرا: 1جدول
850bp F: 5GGTTAAAAAATCACTGCGTCʹ 3 R: 5TTGGTGACGATTTTAGCCGCʹ 3
CTXM-1
تر شود ح شکل کامليتوض CTX-M-1: يژن مورد بررس PCRجه ي: نت2شکل
تر شود ح شکل کامليتوض مثبتESBLs مولدين فنوتيپي تشخيص: 1شکل
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زوله ها. يا يکيوتيب يدرصد مقاومت آنتنمودار : 1نمودار
بحث
Enterobacteriaceae
Enterobacteriaceae ESBL
CTX-M-1PCR
ESBL
SHVTEM CTX-M
blaCTX-M
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.
ESBL
CTX-M-1
CTX-M
E. coli
ESBLs
CTX-M-1
E.coli
MIC
PCR blaCTX-M
blaSHV blaTEM
CTX-M
E. coli
DADPCR ESBL
TEM
160 E.coli
CTX-M PCR
CTX-M-1 CTX-M-5
CTX
CTX-M
SHVTEM
CTX-M-3
PCR
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يريگ جهينت
ESBL
ESBL
تقدير و تشکر
IR.YUMS.REC.139494
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REFERENCES 1.Khalili M B, Sharifi Yazdi M K , Ebadi M, Sadeh M. Correlation between urine analysis and urine culture in the diagnosis of urinary tract infection in Yazd central laboratory. Tehran University Medical Journal TUMS Publications 2007; 65(9): 53-8. 2.Crude N, Tveten Y, Kristiansen BE. Urinary tract infections in Norway: bacterial etiology and susceptibility. A retrospective study of clinical isolates. Clinical Microbiology and Infection 2001; 7(10): 543-7. 3.Amjad A, Mirza IA, Abbasi SA, Farwa U, Sattar A, Qureshi ZA. Spectrum and antimicrobial susceptibility pattern of pathogens causing urinary tract infection-experience in a tertiary care setting. Infectious Disease Journal of Pakistan 2011; 20(2): 56. 4.Farrell D, Morrissey I, De Rubeis D, Robbins M, Felmingham D. A UK multicentre study of the antimicrobial susceptibility of bacterial pathogens causing urinary tract infection. Journal of Infection 2003; 46(2): 94-100 . 5.Zilevica A, Paberza R. Etiological agents of nosocomial urinary tract infections. Proc of LU (LU Zinātniskie Raksti 2005; 3: 63 - 67 6.Li Q, Lee JY, Castillo R, Hixon MS, Pujol C, Doppalapudi VR, et al. NB2001, a novel antibacterial agent with broad-spectrum activity and enhanced potency against β-lactamase-producing strains. Antimicrobial Agents and Chemotherapy 2002; 46(5): 1262-8. 7.Liu G, Ling BD, Zeng Y, Lin L, Xie YE, Lei J. Molecular characterization of extendedspectrum betalactamases produced by clinical isolates of Enterobacter cloacae from a teaching hospital in china. Jpn J Infect Dis 2008; 61(4): 286 . 8.Bush K, Jacoby GA, Medeiros AA. A functional classification scheme for beta-lactamases and its correlation with molecular structure. Antimicrobial Agents and Chemotherapy 1995; 39(6): 1211 9.Thomson K, Prevan A, Sanders C. Novel plasmid-mediated beta-lactamases in enterobacteriaceae: emerging problems for new beta-lactam antibiotics. Current Clinical Topics in Infectious Diseases 1996; 16:, 151 . 10..Shenoy S, Yadav T. The antibiotic susceptibility patterns of uropathogenic Escherichia coli, with special reference to the fluoroquinolones. Journal of Clinical and Diagnostic Research: JCDR 2013; 7(6): 1027 . 11.Vellinga A, Tansey S, Hanahoe B, Bennett K, Murphy AW, Cormican M. Trimethoprim and ciprofloxacin resistance and prescribing in urinary tract infection associated with Escherichia coli: a multilevel model. Journal of Antimicrobial Chemotherapy 2012; 67(10):2523-30 12.Po-Ren H, Wen-Chien KO, JongWu J, JihLu J, DerWang FU. Consensus statement on the adherence to Clinical and Laboratory Standards Institute (CLSI) Antimicrobial Susceptibility Testing Guidelines (CLSI-2010 and CLSI-2010-update) for Enterobacteriaceae in clinical microbiology laboratories in Taiwan. Journal of Microbiology, Immunology and Infection 2010; 43(5): 452-5. 13.Fu Y, Zhang W, Wang H, Zhao S, Chen Y, Meng F, et al. Specific patterns of gyr A mutations determine the resistance difference to ciprofloxacin and levofloxacin in Klebsiella pneumoniae and Escherichia coli. BMC Infectious Diseases 2013; 13(1): 1. 14.Heisig P. Genetic evidence for a role of parC mutations in development of high-level fluoroquinolone resistance in Escherichia coli. Antimicrobial Gents and Chemotherapy 1996: 40(4): 879-85. 15. Watts JL, Jeffrey L., RM(AAM) Thomas R. Shryock, Michael Apley, DVM, Donald J. Bade Steven D. Brown Jeffrey T. Gray, Henry Heine, Rob P. Hunter, MS, Dik J. Mevius, DVM, Mark G. Papich, DVM, Peter Silley, Gary E. Zurenko. Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals; approved standard—third edition. 2008 16 Park YJ, Yu JK, Park KG, Park YG, Lee S, Kim SY, Jeong SH. Prevalence and contributing factors of nonsusceptibility to imipenem or meropenem in extended-spectrum β-lactamase–producing Klebsiella pneumoniae and Escherichia coli. Diagnostic Microbiology and Iinfectious Disease 2011; 71(1): 87-9. 17.Paterson DL, BonomoRA. Extended-spectrum β-lactamases: a clinical update. Clinical Microbiology Reviews 2005; 18(4): 657-86. 18.Cantón R, Novais A, Valverde A, Machado E, Peixe L, Baquero F, & Coque T. Prevalence and spread of extended-spectrum β-lactamase-producing Enterobacteriaceae in Europe. Clinical Microbiology and Infection 2008; 14(s1): 144-53 .
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from
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agha
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at 1
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Feb
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y 11
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19.Shahcheraghi F, Nasiri S, Noveiri H. The survey of genes encoding beta-lactamases, in escherichia coli resistant to beta-lactam and non-beta-lactam antibiotics. Iranian Journal of Basic Medical Sciences 2007; 13(1): 230-7 20.Dallal M, Sabbaghi A, Fallah J, Aghamirzaei HM, Lari A, Eshraghian MR, et al. Evaluation of presence of the bla-SHV and bla-AmpC (CITM, FOX) β-lactamase genes in clinical isolates of Escherichia coli. Journal of Medical Council of Islamic Republic of Iran 2010; 28(3): Pe269-76. 21.Ho PL, Wong R, Chow KH, Yip K, Wong S, Que TL. CTX-M type beta-lactamases among fecal Escherichia coli and Klebsiella pneumoniae isolates in non-hospitalized children and adults. Journal of Microbiology 2008; 41(5): 428-32 . 22.Jonas Bonnedahl , Mirva Drobni , Michel Gauthier-Clerc, Jorge Hernandez, Susanne Granholm, Yves Kayser, Åsa Melhus, Gunnar Kahlmeter, Jonas Waldenström, Anders Johansson, Björn Olsen. Dissemination of Escherichia coli with CTX-M type ESBL between humans and yellow-legged gulls in the south of France. PloS One 2009; 4(6): 23.Shahcheraghi F, Noveiri H, Nasiri S. Detection of bla TEM and bla SHVgenes among clinical isolates of E. coli from Tehran hospitals. Iranian Journal of Medical Microbiology 2007; 1(3): 1-8. 24.Mirzaee M, Pourmand MR, Chitsaz M, Mansouri S. Antibiotic resistance to third generation cephalosporins due to CTX-M-Type extended-spectrum β-lactamases in clinical isolates of Escherichia coli. Iranian Journal of Public Health 2009; 38(1):10-17
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Armaghane-danesh, Yasuj University of Original Article
Medical Sciences Journal (YUMSJ)
Frequency of CTX-M-1 Gene in
Escherichia coli Isolates of
ESBL-Producing Enzyme in Clinical
Samples by Polymerase Chain Reaction
(PCR) in Yasuj
Khishvand Z1, Hosseini M
2, Khoram Rooz SS
1, Sharifi A
1, Tabatabaei SM
3, Khosravani SAM
1*
1Cellular and Molecular Research Center, yasuj University of Medical Sciences, yasuj, Iran,
2Student
Research Committee, yasuj University of Medical Sciences, yasuj, Iran, 3Department of Genycology, Yasuj
University of Medical Sciences, Yasuj, Iran
Received: 03 Feb 2019 Accepted: 03 Des 2018
Abstract Background & aim: Resistance to β-lactam antibiotics in the clinical isolates, in most cases is caused by β-
lactamase enzymes. In recent years, The incidence of broad-spectrum β-lactamase enzymes (ESBLs) among clinical isolates especially E.coli is greatly increased, since the β-lactamase have several subfamilies, using universal primers designed to detect the following complete families could be useful. β-lactamase producing enzymes (ESBLs) of E. coli has created many problems for patients. β-lactamase CTX-M-1 gene is the cause of resistance. The aim of this study was to evaluate CTX-M-1gene in E.coli. Methods: In this practical study, susceptibility of isolated bacteria to 13 antibiotics were indicated by disk
diffusion method according to CLSI guidelines and strains were analyzed for the presence of widespread β-lactamase enzymes via two-disc synergy method. Thus، the prevalence of CTX-M1 ESBL gene samples were determined using PCR and the data were analyzed using ANOVA. Results: A total of 200 isolates of E.coli were isolated. The presence of CTX-M-1 gene were also isolated
using the PCR method. From 200 strains studied, 62 (31%), of strains produced ESBL. After PCR processing of 62 produced ESBL, 43 isolates (69.4%) were identified as CTX-M-1 genes. Also, antibiotic susceptibility test showed the highest percentage of resistance to Cotrimoxazole antibiotic (50%) and the lowest antibiotic resistance to imipenem (0%). Conclusion: The results of this study showed the high percentage of β-lactamase resistance among of E.coli
strains. This is a serious public hazard that should be pointed out to measures for preventing this hazard. Considering the sensitivity of the studied beta-lactam resistant isolates and isolates in Iran to imipenem, a carbapenem with a non-beta-lactam antibiotic is recommended for the treatment of nosocomial infections caused by these strains. Keywords: E.coli, ESBL, PCR, CTXM1
*Corresponding author: Khosravani SAM, Cellular and Molecular Research Center, yasuj University of
Medical Sciences, yasuj, Iran Email: [email protected] Please cite this article as follows: Khishvand Z, Hosseini M, Khoram Rooz SS, Sharifi A, Tabatabaei SM, Khosravani SAM. Frequency of CTX-M-1 Gene in Escherichia Coli Isolates of ESBL-Producing Enzyme in Clinical Samples by Polymerase Chain Reaction (PCR) in Yasuj. Armaghane-danesh 2020; 24(6): 1116-1126.
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