scholarly journals Phenotypic and genotypic detection of extended spectrum β-lactamases among Escherichia coli and Klebsiella pneumoniae isolates from type 2 diabetic patients with urinary tract infections

2021 ◽  
Vol 21 (2) ◽  
pp. 497-504
Author(s):  
Souad Youssouf Kani Elmi ◽  
Medhat Saber Ashour ◽  
Fathy Zakaria Alsewy ◽  
Nashwa Fawzy Abd El Moez Azzam

Background: T2DM patients are more likely to have UTIs caused by resistant organisms such as ESBLs producing bacteria. Challenging reliable identification and prompt characterization of in-vitro susceptibilities of these bacteria are the first steps of deciding the appropriate antimicrobial therapy for UTIs caused by them. Objectives: To isolate and identify E. coli and K. pneumoniae from urine of T2DM patients with UTIs, to determine antibiotic resistance pattern among isolates, and to identify ESBLs production phenotypically and genotypically. Material and method: All samples were cultured on Cystine-Lactose-Electrolyte-Deficient Agar medium (CLED) by using calibrated loop. Growth of 100 colonies or more, i.e. 105 colony forming units (CFU)/mL urine was considered as signifi- cant bacteriuria. Isolation and identification were done according to standard method. All isolates were tested for antibiotic susceptibility testing by the disc diffusion method according to CLSI guidelines. Phenotypic detection of ESBLs was done by double-disk synergy test. Genotypic detection of blaTEM, blaSHV and blaCTX-M genes by using PCR. Results: Results of this study showed that E. coli and K. pneumoniae were the dominant bacterial isolates, they constituted 103 (91.2%) out of 113 urine isolates. E. coli (58. 4%) K. pneumoniae (32.7%), Enterococcus spp. (4.4%), Proteus spp. (2.7%) and Pseu- domonas spp. (1.8%). About 25 (24.3%) out of 103 E. coli and K. pneumoniae isolates were ESBLs positive by DDST, and 22 (88.0%) out of them had ESBLs encoding genes by conventional PCR. The most common gene detected was blaTEM (59.1%), followed by blaSHV (27.3%). CTX-M had not been detected in any of testes isolates. Conclusion: blaTEM and blaSHV genes were detected in 22 out of 25 ESBLs producing E. coli and K. pneumoniae isolates phenotypically detected by DDST. blaTEM was found to be the predominant gene (59.1%), while blaCTX-Mene was not detected in any of tested isolates. Keywords: Extended Spectrum β-Lactamases; Type 2 diabetes mellitus; Urinary tract infections; Phenotypic; genotypic methods.

2017 ◽  
Vol 68 (3) ◽  
pp. 566-569 ◽  
Author(s):  
Mihaela Magdalena Mitache ◽  
Carmen Curutiu ◽  
Elena Rusu ◽  
Ramona Bahna ◽  
Mara Ditu ◽  
...  

One of the most frequent chronic complications occurred in diabetes patients are the urinary tract infections (UTIs). This study aimed to investigate the incidence of UTIs in a cohort of 93 (47 males: 46 females) diabetic patients, the prevalence of different microbial species involved and their virulence and antibiotic resistance profiles. The identification of the uropathogenic strains in the positive urine samples was performed using conventional methods and API tests. After identification, the antibiotic susceptibility profiles were established by the standardized disk diffusion method and double disk diffusion test was performed for the confirmation of ESBL and inducible AmpC b �lactamase phenotypes. The isolated strains were tested for the production of different cell associated and soluble virulence factors, i.e.: bacterial adherence to cellular substrata (HeLa cells), hemolysins (hemolysis spot, CAMP-like), amylase, caseinase, aesculin hydrolysis, DNA-ase, lipase and lecithinase. In the analyzed group, the total prevalence of UTIs was of 46%, a higher incidence being observed in the female patients (64%). Similar to other studies, the etiology of UTI in the investigated diabetes patients was dominated by E. coli, followed by Klebsiella sp. strains. The isolated strains preserved good susceptibility rates to quinolones and aminoglycosides and revealed important virulence features, related to their capacity to colonize the cellular substratum and to produce soluble virulence factors involved in persistence, colonization and progression of the infectious process. The high percentage of beta-lactam resistant strains (including carbapenem-resistant ones) requires careful surveillance of the dynamics of susceptibility profiles for limiting the emergence of these strains in community.


2021 ◽  
Vol 17 (1) ◽  
pp. 119-129
Author(s):  
R.H. Bello ◽  
Y.K.E. Ibrahim ◽  
B.O. Olayinka ◽  
A.A.G. Jimoh ◽  
N.B. Afolabi-Balogun ◽  
...  

Background: The emergence of multidrug- resistance Enterobacteriaceae especially in E. coli bacteria  associated with Urinary Tract Infections (UTIs) in pregnancy is a serious menace globally posing health challenges and confounding successful empirical treatment as well as increasing pregnancy – related  complications.Objectives: The aim of this study is to determine the phenotypic and genotypic characteristics of Extended Spectrum Beta – Lactamases (ESBLs) producing E. coli (ESBLs – EC) isolates in pregnant women attending ante – natal clinics within Ilorin - Kwara State, Nigeria.Materials and methods: A total of 53 non - repeated E. coli isolates from urine samples of pregnant women were presumptively identified using standard bacteriological method and confirmed by commercially available Microgen® Identification Kits. Phenotypic detection of ESBLs was determined using antibiotics susceptibility test and double disc synergy Method for screening and confirmation respectively. Polymerase Chain Reaction (PCR) was further used for the genotypic detection of ESBLs genes.Results: A total 88.67% (47/53) of E. coli exhibited resistance to the cephalosporins of which aztreonam was the highest (75.47%) and the least was cefpodoxime (35.84%) while 85.10% were confirmed positive for ESBL production. The genotypic detection showed the most occurring genotype was blaTEM (50%) blaOXA (27.7%), blaGES (22.5%), blaSHV (15%), blaCTXM and blaVEB (7.5%) while sixty – four (64%) of isolates co – harbored two or more gene. BlaTEM and blaOXA were dominant.Conclusion: This study showed high resistance of E. coli to the third generation cephalosporins harboring different ESBL genes which increases UTIs complexity and limit therapeutic options in pregnancy. Therefore, continuous monitoring of resistance in E. coli, effective appraisal of antibiotic control policies and rational use of antibiotics is therefore encouraged.


2018 ◽  
Vol 12 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Yacoub R. Nairoukh ◽  
Azmi M. Mahafzah ◽  
Amal Irshaid ◽  
Asem A. Shehabi

Background: Emergence of multi-drug resistant uropathogenic E. coli strains is an increasing problem to empirical treatment of urinary tract infections in many countries. This study investigated the magnitude of this problem in Jordan. Methods: A total of 262 E. coli isolates were recovered from urine samples of Jordanian patients which were suspected to have urinary tract infections (UTIs). All isolates were primarily identified by routine biochemical tests and tested for antimicrobial susceptibility by disc diffusion method. Fifty representative Multidrug Resistance (MDR) E. coli isolates to 3 or more antibiotic classes were tested for the presence of resistance genes of blaCTX-M- 1, 9 and 15, carbapenemase (blaIMP, blaVIM, blaNDM-1, blaOXA-48), fluoroquinolones mutated genes (parC and gyrA) and clone of ST131 type using PCR methods. Results: A total of 150/262 (57.3%) of E. coli isolates were MDR. Urine samples of hospitalized patients showed significantly more MDR isolates than outpatients. Fifty representative MDR E. coli isolates indicated the following molecular characteristics: All were positive for mutated parC gene and gyrA and for ST131 clone, and 78% were positive for genes of CTX-M-15, 76% for CTX-M-I and for 8% CTX-M-9, respectively. Additionally, all 50 MDR E. coli isolates were negative for carbapenemase genes (blaIMP, blaVIM, blaNDM-1, blaOXA-48), except of one isolate was positive for blaKPC-2 . Conclusion: This study indicates alarming high rates recovery of MDR uropathogenic E. coli from Jordanian patients associated with high rates of positive ST131 clone, fluoroquinolone resistant and important types of blaCTX-M.


2017 ◽  
Vol 8 (2) ◽  
Author(s):  
Naeimeh Sadat Hashemi ◽  
Meysam Mojiri ◽  
Parivash Yazdani Kachouyi ◽  
Shiva Eskandari ◽  
Mehrsa Mohammadian ◽  
...  

Pseudomonas aeruginosa is one of the most important opportunistic pathogens responsible for various types of hospital infections. High prevalence of antibiotic resistance in P. aeruginosa strains of human clinical samples cause more severe diseases for a longer period of time. The current research was done in order to study the distribution of blaIMP-1 gene among the imipenem-resistant P. aeruginosa strains isolated from burn and urinary tract infections of hospitalized patients. Two-hundred and forty-three P. aeruginosa isolates recovered from the cases of burn and urinary tract infections of inpatients and outpatients were analysis for antibiotic resistance pattern using the disk diffusion method. Then, imipenem-resistant isolates were further analyzed for distribution of blaIMP-1 gene using the PCR. Of 243 P. aeruginosa isolates, 146 strains (60.08%) were taken from outpatients and 97 strains (39.91%) were taken from inpatients. P. aeruginosa isolates harbored the highest levels of resistance against streptomycin (100%), nalidixic acid (100%), aztreonam (100%), cotrimoxazole (95.47%), ciprofloxacin (88.47%), cefotaxime (84.36%) and gentamycin (83.95%). Inpatients had a relatively higher levels of antibiotic resistance. One-hundred and twenty-one out of 126 (96.03%) imipenem-resistant P. aeruginosa isolates harbored the blaIMP-1 gene. Inpatients also had a relatively higher prevalence of blaIMP-1 gene. High prevalence of blaIMP-1 gene and also imipenemresistant P. aeruginosa are important public health issue. Clinical laboratories should consider the detection of the blaIMP-1 gene among the P. aeruginosa isolates of clinical samples.


2013 ◽  
Vol 57 (9) ◽  
pp. 4512-4517 ◽  
Author(s):  
Etienne Ruppé ◽  
Brandusa Lixandru ◽  
Radu Cojocaru ◽  
Çağrı Büke ◽  
Elisabeth Paramythiotou ◽  
...  

ABSTRACTExtended-spectrum-beta-lactamase (ESBL)-producingEscherichia coli(ESBLE. coli) strains are of major concern because few antibiotics remain active against these bacteria. We investigated the association between the fecal relative abundance (RA) of ESBL-producingE. coli(ESBL-RA) and the occurrence of ESBLE. coliurinary tract infections (UTIs). The first stool samples passed after suspicion of UTI from 310 women with subsequently confirmedE. coliUTIs were sampled and tested for ESBL-RA by culture on selective agar. Predictive values of ESBL-RA for ESBLE. coliUTI were analyzed for women who were not exposed to antibiotics when the stool was passed. ESBLE. coliisolates were characterized for ESBL type, phylogroup, relatedness, and virulence factors. The prevalence of ESBLE. colifecal carriage was 20.3%, with ESBLE. coliUTIs being present in 12.3% of the women. The mean ESBL-RA (95% confidence interval [CI]) was 13-fold higher in women exposed to antibiotics at the time of sampling than in those not exposed (14.3% [range, 5.6% to 36.9%] versus 1.1% [range, 0.32% to 3.6%], respectively;P< 0.001) and 18-fold higher in women with ESBLE. coliUTI than in those with anotherE. coliUTI (10.0% [range, 0.54% to 100%] versus 0.56% [range, 0.15% to 2.1%[, respectively;P< 0.05). An ESBL-RA of <0.1% was 100% predictive of a non-ESBLE. coliUTI. ESBL type, phylogroup, relatedness, and virulence factors were not found to be associated with ESBL-RA. In conclusion, ESBL-RA was linked to the occurrence of ESBLE. coliUTI in women who were not exposed to antibiotics and who had the same clone ofE. coliin urine samples and fecal samples. Especially, a low ESBL-RA appeared to be associated with a low risk of ESBLE. coliinfection.


2014 ◽  
Vol 8 (07) ◽  
pp. 818-822 ◽  
Author(s):  
Farzaneh Firoozeh ◽  
Mohammad Zibaei ◽  
Younes Soleimani-Asl

Introduction: Plasmid-mediated quinolone resistance, which complicates treatment, has been increasingly identified in Escherichia coli isolates worldwide. The purpose of this study was to identify the plasmid-mediated qnrA and qnrB genes among the quinolone-resistant Escherichia coli isolated from urinary tract infections in Iran. Methodology: A total of 140 Escherichia coli isolates were collected between March and October 2012 from urinary tract infections in Khorram Abad, Iran. All isolates were tested for quinoloe resistance using the disk diffusion method. Also, all quinolone-resistant isolates were screened for the presence of the qnrA and qnrB genes by polymerase chain reaction. Minimum inhibitory concentrations (MICs) of ciprofloxacin for the qnr-positive isolates were determined. Results: One hundred sixteen (82.8%) of 140 Escherichia coli isolates were nalidixic acid-resistant; among them, 14 (12.1%) and 9 (7.8%) were qnrA and qnrB-positive, respectively. Two quinolone-resistant isolates harbored both qnrA and qnrB. Among 63 ciprofloxacin-resistant isolates, 14 (22.2%) and 9 (14.3%) were found to carry qnrA and qnrB genes, respectively. The ciprofloxacin MIC range was 0.25–512 μg/mL for 23 qnr-positive Escherichia coli isolates, 18 of which had MICs values of 4–512 μg/mL. Conclusion: Our study shows that the frequency of plasmid-mediated quinolone resistance genes among E. coli isolates in Iran is high.


2007 ◽  
Vol 1 (03) ◽  
pp. 257-262 ◽  
Author(s):  
Samuel Kariuki ◽  
Gunturu Revathi ◽  
John Corkill ◽  
John Kiiru ◽  
Joyce Mwituria ◽  
...  

Background: Uropathogenic Escherichia coli are increasingly becoming resistant to flouroquinolones and to other commonly available antimicrobials. We sought to investigate the genetic basis for fluoroquinolone and extended spectrum beta-lactam (ESBL) resistance in 17 fluoroquinolone-resistant (MIC of levofloxacin and ciprofloxacin >32 μg/ml) E. coli isolated from patients with urinary tract infections (UTIs). Methods: We applied PCR and Pulsed Field Gel Electrophoresis (PFGE) to characterize resistance genes and to determine clonal relatedness of strains, respectively. Results: Twelve of the 17 E. coli were resistant to multiple drugs, including ampicillin, co-amoxyclav, cefotaxime, ceftriaxone, ceftazidime and gentamicin and nalidixic acid and produced plasmid-mediated CTX-M-15 type ESBLs and CMY-2 AmpC type enzymes. The other 5 E. coli that were non-ESBL-producing were multiply resistant to ampicillin, nitrofurantoin, cefoxitin, nalidixic acid. Resistance to fluoroquinolones resulted from a combination of the presence of qnrA, qnrB, ciprofloxacin acetylating enzyme designated aac(6’)-1b-cr, and mutations in the two amino acid substitutions; 83 Serine (TCG) to Leucine (TTG) and 87 Aspartic acid (GAC) to Asparagine (AAC). Conclusion: Antibiogram patterns and PFGE of E. coli showed that these were community acquired UTI caused by pockets of clonally-related and some discreet strain types. Plasmid-mediated CTX-M-15 beta-lactamases and CMY-2 AmpC enzymes and fluoroquinolone resistant E. coli are becoming increasingly prevalent in hospitals in Kenya, posing a major challenge in the management of UTIs.


2016 ◽  
Vol 10 (10) ◽  
pp. 1135-1139 ◽  
Author(s):  
Ingrid Cécile Djuikoue ◽  
Paul-Louis Woerther ◽  
Michel Toukam ◽  
Charles Burdet ◽  
Etienne Ruppé ◽  
...  

Introduction: During the last decade, the prevalence of the intestinal carriage of extended spectrum beta-lactamases – producing Escherichia coli (ESBL-E. coli) has continued to increase worldwide in the community, especially in developing countries. Hence, we undertook a study to determine the ESBL-E. coli fecal carriage rate and the associated risk factors in Cameroonian women. Methodology: A total of 86 women suspected of community-acquired urinary tract infections (UTI) were included in 10 health structures from May 2011 to April 2012. After filling a questionnaire, they provided a stool sample that was plated on selective media for ESBL producing bacteria. The identification of strains was obtained with mass spectrometry and the antibiotic susceptibility by disk diffusion in agar media. The ESBL type was determined by PCR. The relative abundance of ESBL-E. coli was measured for positive samples. Eventually, the presence of antibiotics in stool was assessed. Results: The carriage rate of ESBL-E. coli was 57/86 (66.3%). Phenotypic and molecular characterization showed that all ESBL-E. coli strains contained group 1 CTX-M enzymes. Multivariate analysis showed that ESBL-E. coli fecal carriage was associated with the presence of antibiotics in stools (p < 0.05). Although not significant, mean ESBL relative abundance tended to be higher in patients with antibiotic exposure. Conclusions: Our results show that the carriage of ESBL-E. coli fecal carriage in women with UTI suspicion from the Cameroonian community is extremely high and associated with recent antibiotic intake.


2019 ◽  
Vol 70 (11) ◽  
pp. 3788-3792
Author(s):  
Adrian Hasegan ◽  
Maria Totan ◽  
Elisabeta Antonescu ◽  
Adrian Gheorghe Bumbu ◽  
Carmen Pantis ◽  
...  

Urinary tract infections (UTIs) are the most common bacterial pathologies in children, but they are difficult to spot. The diagnosis relies on urine culture in order to measure the prevalence of the infection, to identify the etiology and the sensitivity of the germs to different antibiotics. Escherichia coli (E. coli) strains are the most common uro-pathogen germs. The change in sensitivity to antibiotic of these uro-pathogen bacteria should be closely monitored because the physicians should be informed about the evolution of the antibiotic resistance of E coli, for a more effective treatment in their fight against diseases. The study aimed to determine the prevalence of UTIs and the evolution of antimicrobial sensitivity for E. coli. This retrospective study was performed over a period of 4 years, 2013-2016, and included all the patients admitted in the Children�s Hospital, aged 0-18 years, with the suspicion of UTIs; also, the standard culture techniques for urine samples, the modified Kirby-Bauer disk diffusion method for the antibiotic sensitivity testing, and the disk diffusion method to confirm the ESBL production by the clinical isolates of E. coli in urine were used. The statistical analysis was performed using the proportions of sensitive, resistant and intermediates. Descriptive statistics like the total, mean and percentage were performed using the Statistical Package for the Social Sciences (SPSS), version 15.0 and Microsoft Excel. From 15389 urine cultures processed in 4 years, 1530 were positive (9.9 %). Among these positive urine cultures, 1056 (69 %) were positive for E. coli. Testing the E. coli to a range of antibiotics, according to CLSI standard, a high resistance to Ampicillin (69-96%), Amoxicillin/Clavulanic acid (32-70%), Trimethoprim/Sulfamethoxazole (36-42%) was observed and low levels of resistance to Ceftazidime, Cefuroxime, Cefpodoxime, Gentamycin, Nalidixic acid. Among E. Coli strains, 9-9.6 % were ESBL positive. Despite the low number of positive urine cultures in a paediatric population, it is very important to perform the urine culture in order to correctly identify the etiology of UTIs, recommend the right antibiotic, and avoid the wrong use of the antibiotics in children.


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