scholarly journals Molecular Detection of blaTEM, blaCTX-M and blaSHV Genes in Extended Spectrum β-Lactamase (ESBL) Escherichia Coli from Clinical Samples

Author(s):  
Mahesh Kumar Chaudhary ◽  
Indrani Jadhav ◽  
Megha Raj Banjara

Abstract BackgroundExtended spectrum β-lactamases are the group of beta-lactamase enzymes which confer resistance to the oxyimino-cephalosporins and monobactams. Emergence of ESBL producing genes which possess a serious threat for the treatment of infections both in community and hospitals since it is found to be increasing trends of multidrug resistance. This study was focused to find out the ESBLs producing genes.MethodsThis was a cross-sectional study conducted over a period of 2 years (September 2018 to April 2020) at microbiology laboratory of Nepal Mediciti Hospital. Clinical samples were processed in microbiology laboratory and culture isolates were indetified and characterized by standard microbiological techniques. following standard procedures.Antibiotic susceptibility testing was performed by modified Kirby-Bauer disc diffusion method as recommended by Clinical and Laboratory Standard Institute. Extended spectrum beta-lactamases were phenotypically confirmed by combined disc method.ESBL producing genes i.e. blaTEM,blaCTX-M and blaSHV were confirmed by PCR.ResultsOf the 1449 total E.coli isolates, 323/1449(22.29%) isolates were multi -drug resistance.Among total MDR Escherichia coli isolates, 215/323(66.56%) isolates were ESBL producers. The maximum number of ESBL Escherichia coli was isolated from urine 194(90.23%), followed by sputum 12(5.58%), swab 5 (2.32%), pus 2 (0.93%) and blood 2 (0.93%).Antibiotic susceptibility pattern of ESBL E.coli producers showed highest sensitivity towards tigecycline (100%) followed by polymyxin b, colistin and meropenem.Out of 215 phenotypically confirmed ESBL E.coli,only 186(86.51%) isolates were found to positive by PCR.The last 29(13.49%) were negative for any of the resistant genes.Among the ESBL genotypes,most common was blaTEM 118(63.4%) followed by blaCTX-M 68(36.6%).ConclusionThe emergence of MDR and ESBL producing E.coli isolates with high antibiotic resistant rates to commonly used antibiotics and increased predominance of major gene types blaTEM is a serious concern to the clinicians as well as microbiologist. This study forwarded a real massage to all the clinicians for the emergence of XDR and PDR resistant bacteria and preservation of antibiotics for their proper use in near future, if past experience with MDR and ESBLs is any indicator.

F1000Research ◽  
2018 ◽  
Vol 7 ◽  
pp. 1157 ◽  
Author(s):  
Ahmed Abduljabbar Jaloob Aljanaby ◽  
Israa Abduljabbar Jaloob Aljanaby

Background: Burn infections are one of the most common serious illnesses caused by pathogens, mainly by both gram-negative and gram-positive bacteria. The aim of this study was to detect of the prevalence of multi-drug resistant and extended-spectrum β-lactamase-producing (ESBL) bacteria isolated from inpatients with burn infection and the antimicrobials sensitivity patterns of all bacterial isolates during three years. Methods: This cross-sectional study was performed in Al-Najaf Central Hospital in Al-Najaf City, Iraq from January 2015 to December 2017. A total of 295 burns swabs were collected from hospitalized patients with burn infection. All grown bacterial isolates were identified by standardized microbiological tests. Antimicrobials susceptibility testing was done using the disc diffusion method. Multi-drug, extensive-drug and pan-drug resistant bacteria and extended-spectrum β-lactamase-producing bacteria were determined according to standardized methods and guidelines. Results: Of the 295 burn swabs, 513 different bacteria strains were isolated. Pseudomonas aeruginosa was the most common bacteria with 142 isolates (27.6%) followed by methicillin resistance Staphylococcus aureus 106 isolates (20.6%), while Staphylococcus typhi was the least common bacteria with only 17 isolates (3.3%). 323 (63%) different bacterial strains were isolated from patients who stayed in hospital for 15 days. Most bacterial isolates were resistant to most antimicrobials with high percentages. Out of the 513 bacterial isolates; only 33 isolates (6.4%) were resistant to imipenem 10µg and 464 isolates (90.4%) were multi-drug resistant, 20 isolates (14%) were extensive-drug resistant and 17 isolates (3.3%) were pan-drug resistant. Pseudomonas aeruginosa was the most common ESBL-producing bacteria (51 isolates-35.9%). Conclusions: There was a high prevalence of multi-drug resistant bacteria in burn infection in Al-Najaf hospital. Pseudomonas aeruginosa was the most common multi-drug resistant bacteria, and the most common of ESBL bacteria causing burn infection over the three years.


F1000Research ◽  
2017 ◽  
Vol 6 ◽  
pp. 2103 ◽  
Author(s):  
Sushmita Roy ◽  
Mejbah Uddin Ahmed ◽  
Bhuiyan Mohammad Mahtab Uddin ◽  
Zubair Ahmed Ratan ◽  
Monali Rajawat ◽  
...  

Introduction: Infections due to antibiotic resistant bacteria have increased alarmingly in both developed and developing countries. Unrestrained and rapidly spreading bacterial growth has turned the management of wound infections into a serious challenge. This study aimed to determine the prevalence of different bacterial pathogens and their antibiotic susceptibility in various types of wound infections. Methods:  A cross-sectional study was conducted to collect 105 wound swabs. All isolated bacteria were identified based on colony characteristics, gram stain and standard biochemical tests, and antibiotic susceptibility testing (AST) with the disc diffusion method. Descriptive statistics were used to present the study findings, and all analyses were performed using Stata Version 13. Results:  The rate of isolation of bacteria was 92.3%. Staphylococcus aureus was found to be the most frequent isolate (55.7%), followed by Escherichia coli (23.7%), Pseudomonas spp. (8.2%), and Streptococcus pyogenes (7.2%). Gram-positive bacteria were mostly (60%) found sensitive to vancomycin, azithromycin, gentamicin, imipenem, cefixime, and ceftriaxone in this study. Among the Gram-negative bacteria, Escherichia coli (>60%) showed sensitivity to cefixime, azithromycin, cefuroxime, ceftriaxone, cefotaxime, gentamycin, and ceftazidime. Conclusions: The diversity of isolated bacteria and their susceptibility patterns signify a need to implement a proper infection control strategy, which can be achieved by carrying out antibiotic sensitivity tests of the isolates.


2020 ◽  
Vol 14 (4) ◽  
pp. 127
Author(s):  
Tonny Loho ◽  
Agus Rizal A. H. Hamid ◽  
Rainy Umbas ◽  
Na'imatul Mahanani

Background: Fluoroquinolone has been routinely given as a prophylactic antibiotic to patients undergoing transrectal ultrasound (TRUS)-guided prostate biopsy. Currently, there is no data on the profile of fluoroquinolone-resistant bacteria, its association with post-biopsy sepsis, and alternative prophylactic antibiotics.Methods: A cross-sectional study was conducted in patients undergoing TRUS-guided prostate biopsy at Urology Clinic, Cipto Mangunkusumo National Central General Hospital between August and December 2015. Specimens were taken from rectal swabs prior to biopsy. Gramnegative bacilli and Gram-positive cocci were characterized on the Vitek®2 using GN and GP cards (BioMérieux, USA). Antimicrobial susceptibility testing was done by Kirby Bauer disc diffusion method. Post-biopsy infection was monitored within one week after biopsy by telephone call and diagnosed by clinical examination.Results: A total of 52 patients aged 52-80 years were enrolled. Sixty-six isolates grew from 52 swabs; 14 swabs among them grew 2 isolates. The commonest pathogen was Escherichia coli (78.8%), followed by Klebsiella pneumoniae (13.6%), Enterococcus faecium (3.0%), Acinetobacter haemolyticus, Morganella morganii subsp. morganii, and Enterococcus faecalis (1.5%), respectively. Ciprofloxacin-resistant bacteria were found in 33 (50.0%) isolates which were predominated by E. coli. Meanwhile, 90.4% and 96.2% of isolates were sensitive to amoxicillinclavulanate and ampicillin-sulbactam, respectively. Post-biopsy infection was established in 7 patients requiring no hospitalization.Conclusions: The proportion of ciprofloxacin-resistant bacteria in the rectum among patients undergoing prostate biopsy was 50.0%. Escherichia coli is the commonest resistant pathogen to fluoroquinolone. The recommended alternatives for prophylactic antibiotics are amoxicillinclavulanate and ampicillin-sulbactam. Routine pre-biopsy rectal swab cultures should also be encouraged.


2020 ◽  
Vol 3 (2) ◽  
pp. 102-110
Author(s):  
Mahesh Kumar Chaudhary ◽  
Indrani Jadhav ◽  
Indrani Jadhav ◽  
Megha Raj Banjara

Background: Extended spectrum β-lactamases have addressed the serious challenges worldwide due to the emergence of ESBL producing genes which possess a serious threat for the treatment of infections both in community and hospitals since it is found to be increasing trends of multidrug resistance. This study was focused to find out the antibiotic profile of multidrug resistant Escherichia .coli and status of ESBLs producing E.coli. Methods: This was a cross-sectional study conducted over a period of 2 years (September 2017 to April 2019) at microbiology laboratory of Nepal Mediciti Hospital. A total of 16542 samples were processed. Various clinical samples were collected from both inpatients and outpatients aseptically and without contaminating skin commensals. Standard microbiological techniques were used for isolation and identification of pathogens. Extended spectrum beta-lactamases were phenotypically confirmed by combined disc method. Results: Out of 1449 E.coli isolates, 323(22.29%) were found to be MDR E.coli. Isolation rate of ESBL producing E.coli (66.56%) were found to be high among MDR E.coli isolates. Conclusion: There was increasing prevalence of ESBL producing E.coli and was essential to monitor antibiotic susceptibility pattern and formulate antibiotic policy to prevent the spread of MDR and ESBL producers.  


2015 ◽  
Vol 8 (1) ◽  
pp. 71-75 ◽  
Author(s):  
Olga I Chub ◽  
Aleksandr V Bilchenko ◽  
Igor Khalin

Background : Increased multidrug resistance of extended-spectrum beta-lactamases (ESBLs) compromises the efficacy of treatment of urinary tract infections. Objective : The objective of this study is to determine the prevalence of ESBL-producing uropathogens from hospitalized patients with chronic pyelonephritis and to identify the presence of genes involved in the resistance. Methods : A cross-sectional study of 105 patients with chronic pyelonephritis, treated in Kharkiv City Clinical Emergency Hospital, Ukraine was carried. Bacterial isolates were collected, antimicrobial susceptibility of isolates was determined by the Kirby Bauer disk diffusion method and screening for the presence of blaSHV, blaTEM, blaCTX-M ESBL genes was performed by polymerase chain reaction. Results : 84 (80%) patients had positive urine cultures. Eschеrichia coli wаs the most common microorganism isolated. Among them, 29 (25.2%) were found to be ESBL producers. Out of 53 E. coli isolates, 10 (18.9%), 4 (7.5%) and 6 (11.3%) were identified to carry bla(TEM), bla(SHV) and bla(CTX-M) beta-lactamase genes, respectively. The highest resistance was observed against ampicillin (75.9%), ciprofloxacin (48.3%), levofloxacin (41.4%) and gentamicin (41.4%). Beside this, only meropenem (96.6% susceptibility), nitroxolinum (86.2%) and fosfomycin (72.4%) exhibited a good enough activity against ESBLs-producing urinary strains. Conclusion : Isоlation and detеction of ESBL-prоducing strаins are еssential fоr the sеlection оf the mоst effеctive antibiоtic for the empiric trеatment.


2020 ◽  
Vol 13 (1) ◽  
pp. 23-31
Author(s):  
Binita Koirala Sharma

Background: Hands of healthcare workers (HCWs) could be colonized by potential drug resistant bacteria like Extended Spectrum Beta Lactamase producers (ESBLs) and Carbapenems-resistant (CR) isolates and could become vectors of nosocomial pathogens in healthcare facilities that are associated with an increase of morbidity, mortality and healthcare costs. This study aimed to investigate the prevalence of ESBLs and CR isolates from hands of HCWs with their antibiotic susceptibility pattern. Materials and methods: This was a cross-sectional study that included a total of 150 hand swabs collected from March, 2018 to September, 2018 in Gandaki Medical College and Teaching Hospital. Isolation, identification and antimicrobial susceptibility tests were done using standard microbiological procedures. Results: Among the total isolates of 219 obtained from growth positive samples 92/219(42.01%) were Gram negative bacteria (GNB) and the most common were Klebsiella spp 32(34.78%) followed by Escherichia coli 17(18.48%), Pseudomonas aeruginosa12 (13.04%), Acenetobacter spp 11(11.96%), Proteus spp 9(9.78%), Citrobacter spp 7(7.61%) and Enterobacter spp 4(4.35%). The prevalence of ESBLs, CR and ESBLs with Co-resistant to Carbapenems were 19.56%,14.13% and 9.78%, respectively. The most effective drugs for isolates were Nitrofurantoin followed by Amikacin, Tetracycline and Gentamycin. Distribution pattern of the ESBLs and CR isolates among doctors, nurses, laboratory technicians, helpers and basic science faculties were not significant (p>0.05). Conclusions: This report revealed the emerging and moderately high prevalence of ESBLs, CR and ESBLs with Co-resistant to Carbapenems GNB with their antibiotic susceptibility patterns found on hands of HCWs in Nepal. Thus, this study could be helpful in developing proper guidelines on hand hygiene and implementation of infection control measures including contact precautions against the spread of infections by such pathogens in healthcare settings.  


2018 ◽  
Vol 2018 ◽  
pp. 1-7 ◽  
Author(s):  
Teshale Worku ◽  
Dejene Derseh ◽  
Abera Kumalo

Background. Nosocomial infections occur among patients during their stay in hospitals. The severity of infection depends on the characteristics of microorganisms with a high risk of being acquired when the environment is contaminated. Antibiotic-resistant bacteria are emerging rapidly around the globe creating a serious threat. Methods. A cross-sectional study was conducted from December 2016–February 2017 at Mizan-Tepi University Teaching Hospital, Southwest Ethiopia. Samples were collected from the equipment and hospital surfaces. The isolated bacteria were checked for susceptibility by the Kirby–Bauer disc diffusion method following the standards of CLSI 2014. Health professionals and sanitary team members were included in the study which assessed the disinfection practice of objects from which samples were taken. Data were analyzed using SPSS version 20.0. Results. A total of 201 swab samples were taken, and most bacteria were recovered from thermometer and floor consisting of 21.6% S. aureus, 19.3% CoNS, 15.9% E. coli, 14.8% Klebsiella species, 11.4% P. aeruginosa, 10.2% Proteus species, and 6.8% Serratia species. The most multidrug resistant organisms were S. aureus (79%), Klebsiella species (53.8%), CoNS (47%), and Proteus species (44.4%). Only 6.45% of health professionals disinfect their stethoscope consistently. Conclusion. S. aureus, CoNS, and E. coli were the predominant isolates. Most isolates showed highest susceptibility to ciprofloxacin and least to ampicillin and penicillin. There is no regular sanitation and disinfection of hospital equipment and surfaces.


2015 ◽  
Vol 78 (1) ◽  
pp. 72-77 ◽  
Author(s):  
ALENA SKOČKOVÁ ◽  
KATEŘINA BOGDANOVIČOVÁ ◽  
IVANA KOLÁČKOVÁ ◽  
RENÁTA KARPÍŠKOVÁ

The occurrence of antimicrobial-resistant bacteria is an important public health issue. The aim of this study was the monitoring of resistant Escherichia coli in raw cow's milk with a focus on the detection of extended-spectrum β-lactamase (ESBL)–producing strains. In total, 263 samples of raw milk from 40 farms were collected and investigated in 2010 to 2013 in the Czech Republic. Detection of E. coli was performed and evaluated according to ISO 16649-2, and antibiotic resistance was screened by the disk diffusion method. The presence of E. coli was detected in 243 (92.4%) samples. In total, 270 isolates were obtained. Resistance to β-lactam (31.8%) and tetracycline (13.0%) antibiotics was detected most often and also multiresistant strains (5.5%) were observed. E. coli isolates found to be resistant to β-lactam, tetracycline, and quinolone antibiotics were assayed by PCR to detect selected genes encoding those resistance mechanisms. In isolates in which any bla genes were detected, a double-disk synergy test was performed. ESBL production was confirmed in 2 (0.7%) isolates. The genetic analysis identified the presence of the blaCTX-M gene and other resistance genes (tet(B) and qnrB). Both ESBL-positive isolates originated from the same farm and had an identical pulsed-field gel electrophoresis profile. The findings of our study indicate that milk can be a reservoir of bacteria carrying resistance genes with a potential for spreading through the food chain.


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