scholarly journals Molecular detection of β-lactamase genes in Klebsiella pneumoniae and Escherichia coli isolated from different clinical sources

2022 ◽  
Vol 67 (4) ◽  
pp. 170-180
Author(s):  
Kamal Ismael Bakr ◽  
Sherko Muhammed Abdul-Rahman ◽  
Rebwar Muhammad Hamasalih
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Molecular Detection ◽  
Multidrug Resistant ◽  
Clinical Samples ◽  
Chain Reaction ◽  
E Coli ◽  
Recent Emergence ◽  
Polymerase Chain ◽  
Pcr Test

The rising occurrence of infections generated by Escherichia coli and Klebsiella pneumoniae that produce extended-spectrum β-lactamase (ESBL) is reason for concern. Due to the recent emergence of multidrug-resistant microorganisms that develop ESBL. The purpose of this work was to detect the ESBLs in clinical isolates of E. coli and K. pneumoniae. 118 samples of E. coli and 63 isolates of K. pneumoniae were collected from clinical samples. Polymerase chain reaction was used to detect β-lactamase genes (i.e., blaTEM, blaSHV, and blaCTX-M). Phenotypic detection revealed that 48.31% and 85.19% of E. coli and K. pneumoniae produced ESBLs, respectively. Whereas screening of ESBL genes in both bacteria employing a multiplex PCR test revealed that 24.58% of the ESBL-producing E. coli strains contained blaTEM, 50.85% contained blaSHV, and 32.2% contained blaCTX-M. Nevertheless, in K. pneumoniae, 40.74% blaTEM, 35.19% blaSHV, and 64.81% blaCTX-M genes were present. Antimicrobial resistance profiles of E. coli and K. pneumoniae isolates to twenty antibiotics were observed to vary significantly. Additionally, it was determined that the majority of E. coli and K. pneumoniae isolates were multidrug resistant (MDR). Additionally, 80.51% of E. coli isolates were resistant to the AMC antibiotic, while 0.00% were resistant to IPM and MEM. From the other hand, the resistant proportion of K. pneumoniae isolates was heterogeneous, ranging from 69.84% against CAZ to 0.00% against CIP and G antibiotics. The blaSHV gene was the most widespread among different forms of ESBLs in E. coli, but the most common gene in K. pneumoniae isolates was blaCTX-M (64.81%).

scholarly journals Prevalence and molecular detection of shiga toxin producing Escherichia coli from diarrheic cattle

2016 ◽  
Vol 14 (1) ◽  
pp. 63-68 ◽  
Author(s):  
MM Akter ◽  
S Majumder ◽  
KH MNH Nazir ◽  
M Rahman
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Molecular Detection ◽  
Chain Reaction ◽  
Specific Primers ◽  
Fecal Samples ◽  
E Coli ◽  
Uremic Syndrome ◽  
Polymerase Chain ◽  
Positive Isolate

Shiga toxin-producing Escherichia coli (STEC) are zoonotically important pathogen which causes hemorrhagic colitis, diarrhea, and hemolytic uremic syndrome in animals and humans. The present study was designed to isolate and identify the STEC from fecal samples of diarrheic cattle. A total of 35 diarrheic fecal samples were collected from Bangladesh Agricultural University (BAU) Veterinary Teaching Hospital. The samples were primarily examined for the detection of E. coli by cultural, morphological and biochemical characteristics, followed by confirmation of the isolates by Polymerase Chain Reaction (PCR) using gene specific primers. Later, the STEC were identified among the isolated E. coli through detection of Stx-1 and Stx-2 genes using duplex PCR. Out of 35 samples, 25 (71.43%) isolates were confirmed to be associated with E. coli, of which only 7 (28%) isolates were shiga toxin producers, and all of them were positive for Stx-1. However, no Stx-2 positive isolate could be detected. From this study, it may be concluded that cattle can act as a reservoir of STEC which may transmit to human or other animals.J. Bangladesh Agril. Univ. 14(1): 63-68, June 2016

scholarly journals Molecular Detection of Plasmid-Mediated Quinolone Resistance in Ciprofloxacin-Resistant Escherichia coli from Urine of Patients attending Garki Hospital, Abuja, Nigeria

2020 ◽  
Vol 1 (4) ◽  
Author(s):  
Moses Oghenaigah Eghieye ◽  
Istifanus Haruna Nkene ◽  
Rejoice Helma Abimiku ◽  
Yakubu Boyi Ngwai ◽  
Ibrahim Yahaya ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Urinary Tract ◽  
Molecular Detection ◽  
Quinolone Resistance ◽  
Chain Reaction ◽  
E Coli ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Tract Infections

Urinary tract infections (UTIs) caused by Escherichia coli (E. coli) is common worldwide; and its successful treatment using antibiotics is limited by acquisition of resistance by the bacteria. This study investigated the occurrence of plasmid-mediated quinolone resistance (PMQR) genes in ciprofloxacin-resistant E. coli from urine of patients with suspected cases of UTIs attending Garki Hospital Abuja (GHA), Nigeria. A total of 8 confirmed ciprofloxacin-resistant E. coli was screened for carriage of PMQR genes using polymerase chain reaction (PCR) method. The occurrences of the PMQR genes detected were in the order: aac-(6′)-Ib-cr (87.5%) > qnrB (50.0%) > qnrS (37.5%) > oqxAB (12.5%) > qnrA(0.0%). qnrB and qnrS did not exist alone, but in combination with other genes; aac-(6′)-Ib-crexisted both alone and in combination with others; the most prevalent patterns of existence were aac-(6′)-Ib-cr alone and aac-(6′)-Ib-cr + qnrB + qnrS at 25.0% each. This study has shown that the ciprofloxacin-resistant E. coli harbored aac-(6′)-Ib-cr, qnrB, qnrS and oqxAB PMQR genes, with aac-(6′)-Ib-cr being the most prevalent. The genes were present either alone or in combination with one another. This has implication for the clinical application of fluoroquinolones to treat UTI in the study location and environs. 

scholarly journals Phenotypic and Molecular Detection of Resistance Genes from Multi-drug Resistant Escherichia coli Isolated from Patients Attending Selected Hospitals in Damaturu, Yobe State, Nigeria

2021 ◽  
Vol 8 (9) ◽  
pp. 396-407
Author(s):  
Sheriff Wakil ◽  
Mustafa Alhaji Isa ◽  
Adam Mustapa
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Resistance Genes ◽  
Molecular Detection ◽  
Multidrug Resistant ◽  
Clinical Samples ◽  
E Coli ◽  
Resistant Genes ◽  
Tract Infections

Multidrug resistance among Escherichia coli causing urinary tract infections (UTIs) and diarrhea are major public health problem worldwide which cause difficulty in treating the infections caused by Escherichia coli due to the high resistances. The study is aimed to determine the phenotypic and molecular detection of multidrug resistant E. coli isolated from clinical samples of patients attending selected Hospitals in Damaturu, Yobe State-Nigeria. Methods: Two hundred (200) clinical samples were collected aseptically from patient diagnosed with (100 stool samples) and UTI’s (100 urine samples) using sterile universal container. The samples were processed using standard microbiological methods for identification of E. coli. Samples were cultured on MacConkey agar (stool) and Cystine lactose electrolyte deficient agar (urine). The resulting colonies of isolates were further subculture on Eosin methylene blue agar for confirmatory and followed by gram stain, biochemical identification at Microbiology laboratory unit of Yobe State Specialist and Yobe State Teaching Hospital respectively. The antimicrobial susceptibility patterns were determined using Kirby-Bauer disc diffusion techniques and the phenotypic expression of extended spectrum beta-lactamases (ESBLs) were determined using modified double disc synergy test (MDDST) and also the three (3) resistance genes (blaTEM, accC1 and qnrA) were detected using polymerase chain reaction. Results: One hundred and twenty-two (122) isolates were resistant to antibiotics. The highest level of resistance was against amoxicillin (90.2%) while the least resistance was against sparfloxacin (24.3%). Thirty-seven (37) E. coli isolates shows MDR; the highest MDR was (24.3%) while least MDR was (5.4%). The PCR amplification of resistant genes (blaTEM, accC1 and qnrA) were detected on E. coli that shows positive ESBL and the bands were separated using agarose gel electrophoresis. Conclusion: The findings of this study show augmentin, ciprofloxacin and sparfloxacin are the most effective antibiotics against E. coli isolated from patients attending the two hospitals in Damaturu; who are diagnose with UTI and diarrheic infection. The resistant genes include; blaTEM, accC1 and qnrA coding for beta-lactam, aminoglycoside and quinolones were present in E. coli isolated from patients attending selected Hospitals in Yobe State, Nigeria. Keywords: Multidrug resistant, Escherichia coli, extended spectrum beta lactamase, resistance-associated genes, urinary tract infections, diarrheic.

scholarly journals Screening of AmpC-/ESBL-producing Escherichia coli isolates from livestock for STEC/EHEC virulence genes

2021 ◽  
Vol 72 (3) ◽  
pp. 3147
Author(s):  
F PEHLIVANOGLU
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Virulence Genes ◽  
Multidrug Resistant ◽  
Beta Lactamase ◽  
Chain Reaction ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Polymerase Chain ◽  
Flagellar Antigen

Livestock is an important reservoir of Shiga toxin-producing Escherichia coli and enterohemorrhagic E. coli (STEC/EHEC) strains and acts as a significant source of transmission to humans. In addition to the virulence of STEC/EHEC isolates, antibiotic resistance is also an escalating problem in these bacteria and increases the risk to public health. Therefore, the present study aimed to explore E. coli O157:H7 serotype and STEC/EHEC virulence genes in AmpC- and extended-spectrum beta-lactamase (ESBL)-producing E. coli isolates from cattle, chicken and sheep. A total of 61 confirmed AmpC- or ESBL-producing E. coli isolates were screened for the virulence genes (stx1, stx2, eae, ehxA, espP, katP and saa) and E. coli O157 (rfbO157) and H7 (fliCH7) genes by polymerase chain reaction (PCR). None of the ESBL-producing E. coli was positive for these genes, but six multidrug-resistant AmpC-producing E. coli were positive for the fliCH7 gene only. When considering the function of the H7 flagellar antigen of E. coli, it may be concluded that the development of ESBL/AmpC beta-lactamase production in the E. coli isolates with H7 flagella, which reside in the chicken intestine, may be potentially important for public health regarding both virulence and antimicrobial resistance.

scholarly journals Co-Occurrence of Plasmid-Mediated AmpC β-Lactamase Activity Among Klebsiella pneumoniae and Escherichia Coli

2017 ◽  
Vol 11 (1) ◽  
pp. 195-202 ◽  
Author(s):  
Abdulaziz Zorgani ◽  
Hiyam Daw ◽  
Najib Sufya ◽  
Abdullah Bashein ◽  
Omar Elahmer ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Multidrug Resistant ◽  
Control Measures ◽  
Gene Encoding ◽  
E Coli ◽  
Infection Control Measures ◽  
Genes Encoding ◽  
Polymerase Chain ◽  
Variant Enzyme

Introduction: Extended-spectrum β-lactamases (ESBLs), including the AmpC type, are important mechanisms of resistance among Klebsiella pneumoniae and Escherichia coli isolates. Objective: The aim of the study was to investigate the occurrence of AmpC-type β-lactamase producers isolated from two hospitals in Tripoli, Libya. Methods: All clinical isolates (76 K. pneumoniae and 75 E. coli) collected over two years (2013-2014) were evaluated for susceptibility to a panel of antimicrobials and were analyzed phenotypically for the ESBL and AmpC phenotype using E-test and ESBL and AmpC screen disc test. Both ESBL and AmpC-positive isolates were then screened for the presence of genes encoding plasmid-mediated AmpC β-lactamases by polymerase chain reaction (PCR). Results: Of the K. pneumoniae and E. coli tested, 75% and 16% were resistant to gentamicin, 74% and 1.3% to imipenem, 71% and 12% to cefoxitin, 80% and 12% to cefepime, 69% and 22.6% to ciprofloxacin, respectively. None of the E. coli isolates were multidrug resistant compared with K. pneumoniae (65.8%). K. pneumoniae ESBL producers were significantly higher (85.5%) compared with (17.3%) E. coli isolates (P <0.0001, OR=4.93). Plasmid-mediated AmpC genes were detected in 7.9% of K. pneumoniae, and 4% E. coli isolates. There was low agreement between phenotypic and genotypic methods, phenotypic testing underestimated detection of AmpC enzyme and did not correlate well with molecular results. The gene encoding CMY enzyme was the most prevalent (66.6%) of AmpC positive isolates followed by MOX, DHA and EBC. Only one AmpC gene was detected in 5/9 isolates, i.e, blaCMY (n=3), bla MOX (n=1), blaDHA (n=1). However, co-occurrence of AmpC genes were evident in 3/9 isolates with the following distribution: bla CMY and blaEBC (n=1), and blaCMY and blaMOX (n=2). Neither blaFOX nor blaACC was detected in all tested isolates. All AmpC positive strains were resistant to cefoxitin and isolated from patients admitted to intensive care units. Conclusion: Further studies are needed for detection of other AmpC variant enzyme production among such isolates. Continued surveillance and judicious antibiotic usage together with the implementation of efficient infection control measures are absolutely required.

scholarly journals Carbapenemase production in hospital isolates of multidrug-resistant Klebsiella pneumoniae and Escherichia coli in Serbia

2017 ◽  
Vol 74 (8) ◽  
pp. 715-721 ◽  
Author(s):  
Anika Trudic ◽  
Zora Jelesic ◽  
Mira Mihajlovic-Ukropina ◽  
Deana Medic ◽  
Branka Zivlak ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Carbapenem Resistance ◽  
Multidrug Resistant ◽  
Clinical Samples ◽  
Reference Laboratory ◽  
Bacterial Strains ◽  
Beta Lactamases ◽  
E Coli ◽  
Novi Sad

Background/Aim. Carbapenem resistance has escalated in medically important enterobacteria such as Klebsiella pneumoniae and Escherichia coli worldwide. Multidrug-resistant strains represent an important source of concern as effective therapeutic options of infections they cause are limited or none. There were no comprehensive studies considering the presence of carbapenemase production in enterobacteria in Serbia so far. The aim of the study was to determine carbapenemase production in hospital isolates of multidrug-resistant K. pneumoniae and E. coli in Serbia. Methods. Strains of K. pneumoniae and E. coli resistant to at least one carbapenem (imipenem, meropenem, ertapenem) were collected from November 2013 to May 2014. Isolates were obtained from clinical samples of patients treated in 14 hospitals in Serbia. Carbapenem resistance was confirmed using phenotypic tests and polymerase chain reaction (PCR) in National Reference Laboratory for Registration and Surveillance of Antimicrobial Resistance of Bacterial Strains in Novi Sad. Results. Of 129 collected strains, 121 (93.8%) were K. pneumoniae and 8 (6.2%) were E. coli. Seventy (54.3%) strains were obtained from urine, 26 (20.2%) from blood, 19 (14.7%) from wound secretions and 14 (10.9%) from lower respiratory tract secretions. Carbapenemase genes were detected in 58 (45%) isolates. The gene bla New Delhimetallo-beta-lactamases (blaNDM) was found in 33 (27.3%) K. pneumoniae, bla oxacillinases-48 (blaOXA-48) in 10 (8.3%), bla K. pneumonia carbapenemase (blaKPC) in 1 (0.8%), and 7 (5.4%) strains harbored both blaOXA-48 and blaNDM. Seven E.coli harbored blaNDM gene. Conclusions. In Serbia, the most common type of carbapenemase in both multidrug-resistant K. pneumoniae and E. coli is NDM. Co-production of OXA-48 and NDM was found in K. pneumoniae. To our knowledge, KPC production was detected for the first time in Serbia.

scholarly journals Emergence of colistin resistance in multidrug-resistant Klebsiella pneumoniae and Escherichia coli strains isolated from cancer patients

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Mai M. Zafer ◽  
Hadir A. El-Mahallawy ◽  
Asmaa Abdulhak ◽  
Magdy A. Amin ◽  
Mohamed H. Al-Agamy ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Cancer Patients ◽  
Clonal Diversity ◽  
Multidrug Resistant ◽  
Resistance Mechanisms ◽  
Global Public Health ◽  
Colistin Resistance ◽  
E Coli ◽  
Recent Emergence

Abstract Background Colistin resistance is mainly driven by alterations in the Gram-negative outer membrane lipopolysaccharides and is caused, in most cases, by mutations in mgrB gene. However, the recent emergence of plasmid-encoded colistin resistance among Enterobacteriaceae strains represents a serious threat to global public health. In this paper we have investigated the rates of colistin resistance and the underlying mechanisms in 450 Klebsiella pneumoniae and Escherichia coli isolates obtained from cancer patients in Egypt. Methods Colistin susceptibility and minimum inhibitory concentrations were determined according to the European Committee on Antimicrobial Susceptibility Testing, by broth microdilution, and by E-test. The mcr-1, mcr-2 and mgrB genes were detected by PCR and then sequenced. Clonal diversity in colistin-resistant K. pneumoniae was evaluated by multilocus sequence typing. Results Forty (8.8%) colistin-resistant isolates, including 22 K. pneumoniae and 18 E. coli, were isolated over 18 months. Of these, 50% were carbapenem-resistant, out of which nine were blaOXA-48 and seven blaNDM-1 positive. The mechanisms of colistin resistance could be revealed only in three of the 40 resistant strains, being represented by mcr-1 in one blaNDM-1-positive E. coli strain and in one K. pneumoniae ST11 and by mgrB mutations, detected in one K. pneumoniae isolate. None of the studied isolates harbored mcr-2. Conclusions Our results demonstrate a high frequency of colistin resistance in enterobacterial strains isolated from cancer patients, but a low prevalence of the most well known resistance mechanisms.

Molecular Characterization of Antimicrobial Drug Resistance in Escherichia coli Isolated from Clinical Samples

2017 ◽  
Vol 8 (01) ◽  
Author(s):  
Ibtisam Habeeb AL-Azawi ◽  
Aqeel Reheeum Hassan ◽  
Alaa Hamza Jaber
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Antimicrobial Drug Resistance ◽  
Biochemical Characteristics ◽  
E Coli ◽  
Medical City ◽  
Polymerase Chain

A total of 49 different clinical samples (urine n=30, stool n=10, and blood n=8) were collected from patient admitted to the Al-Sadder medical City in Al-Najaf Governorate-Iraq. The results demonstrated that 49 specimens (100%) were diagnosed as E. coli by cultural, biochemical characteristics and Vitek2® system. Polymerase Chain Reaction has been used to detect of some genes which coding antimicrobial resistance in E. coli isolates. Regarding genes that responsible for ESBL enzymes (blaCTX-M, blaOXA and blaTEM), the current results proved that blaTEM genes have highest rate (97.95%) followed by blaTEM and blaOXA (93.75%) for each.

scholarly journals Long-Term Carriage of Ciprofloxacin-Resistant Escherichia coli Isolates in High-Risk Nursing Home Residents

2016 ◽  
Vol 37 (4) ◽  
pp. 440-447 ◽  
Author(s):  
Miriam D. Ismail ◽  
Ting Luo ◽  
Sara McNamara ◽  
Bonnie Lansing ◽  
Evonne Koo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
High Risk ◽  
Polymerase Chain Reaction Amplification ◽  
Nursing Home Residents ◽  
Multidrug Resistant ◽  
Chain Reaction ◽  
E Coli ◽  
Polymerase Chain ◽  
Gram Negative Organisms

BACKGROUNDRates of multidrug-resistant gram-negative organisms are surpassing those of methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci in nursing homes (NHs).OBJECTIVETo characterize the incidence and duration of carriage of ciprofloxacin-resistant Escherichia coli (CipREc) in NHs and identify those in the O25b-ST131 lineage.METHODSWe collected 227 CipREc isolates obtained by routine and regular surveillance of high-risk NH residents with indwelling devices. Repetitive element palindromic (REP)–polymerase chain reaction assay and multiplex polymerase chain reaction amplification for O25b-ST131 E. coli detection were performed using (GTG)5-primers and O25pabBspe and trpA2 primer pairs, respectively.RESULTSWe found a high period prevalence of CipREc colonization (21.5%), high rates of recolonization with the same strain following clearing (0.46 recolonizations/ person/ year), and an acquisition incidence of 1.05 cases/1,000 person-days. Almost three-quarters of colonized residents carried strains in the O25b-ST131 E. coli lineage. Compared with isolates not in the lineage, O25b-ST131 isolates were carried significantly longer (10 vs 3 months). We identified 18 different REP-types; 2 occurred in 55% of the residents colonized with CipREc, and in more than 1 NH. Duration of CipREc carriage varied by REP-type and averaged 6 months.CONCLUSIONCipREc occurred frequently in NH residents and is carried for long durations, and reacquisition following clearance is commonTrial registration. ClinicalTrials.gov identifier: NCT01062841.Infect. Control Hosp. Epidemiol. 2016;37(4):440–447

scholarly journals On-column Refolding of an Insoluble His6-tagged Recombinant EC-SOD Overexpressed in Escherichia coli

2005 ◽  
Vol 37 (4) ◽  
pp. 265-269 ◽  
Author(s):  
Xi-Qiang Zhu ◽  
Su-Xia Li ◽  
Hua-Jun He ◽  
Qin-Sheng Yuan
Keyword(s):  
Escherichia Coli ◽  
Inclusion Bodies ◽  
Mass Ratio ◽  
Chain Reaction ◽  
Expression Plasmid ◽  
Protein Subunits ◽  
E Coli ◽  
Metal Affinity ◽  
Polymerase Chain ◽  
Escherichia Coli Expression

Abstract The EC-SOD cDNA was cloned by polymerase chain reaction (PCR) and inserted into the Escherichia coli expression plasmid pET-28a(+) and transformed into E. coli BL21(DE3). The corresponding protein that was overexpressed as a recombinant His6-tagged EC-SOD was present in the form of inactive inclusion bodies. This structure was first solubilized under denaturant conditions (8.0 M urea). Then, after a capture step using immobilized metal affinity chromatography (IMAC), a gradual refolding of the protein was performed on-column using a linear urea gradient from 8.0 M to 1.5 M in the presence of glutathione (GSH) and oxidized glutathione (GSSG). The mass ratio of GSH to GSSG was 4:1. The purified enzyme was active, showing that at least part of the protein was properly refolded. The protein was made concentrated by ultrafiltration, and then isolated using Sephacryl S-200 HR. There were two protein peaks in the A280 profile. Based on the results of electrophoresis, we concluded that the two fractions were formed by protein subunits of the same mass, and in the fraction where the molecular weight was higher, the dimer was formed through the disulfide bond between subunits. Activities were detected in the two fractions, but the activity of the dimer was much higher than that of the single monomer. The special activities of the two fractions were found to be 3475 U/mg protein and 510 U/mg protein, respectively.

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