Occurrence of ESBLs in Clinical Isolates of Klebsiella Species and Comparative Analysis of Phenotypic Detection Methods

2020 ◽  
Vol 18 (3) ◽  
pp. 255-260
Author(s):  
Taqdees Malik ◽  
Asma Naim
Keyword(s):  
Detection Methods ◽  
Phenotypic Characterization ◽  
Clinical Samples ◽  
Zone Of Inhibition ◽  
Diagnostic Laboratory ◽  
Klebsiella Species ◽  
Beta Lactamases ◽  
Extended Spectrum ◽  
Extended Zone ◽  
Synergy Test

Aims: Drug resistance enzymes like beta-lactamases and Extended-Spectrum Beta- Lactamases (ESBLs) are mostly produced in the response of mutation and a few other factors. These enzymes are plasmid-mediated that can confer resistance against several β-lactam agents including cephalosporin and extended-spectrum cephalosporin. This study is conducted to determine the frequency of ESBL producing Klebsiella strains and to compare the phenotypic detection methods among various clinical samples collected from a commercial centralized Microbiology diagnostic laboratory. Materials and Methods: 125 Klebsiella strains were collected for this study. After identification and susceptibility testing, the phenotypic characterization of ESBL producing K. pneumoniae (n=89) and K. oxytoca (n=36) species was performed by using the double disc synergy test and the combination disc test. Results: In our study, 80 (64%) Klebsiella isolates were found to be ESBL positive due to their ability to produce an extended zone of inhibition with combination disc whereas 45 (36%) isolates were ESBL negative with no extended zone of inhibition. The phenotypic findings obtained by using the combination disc method and double disc synergy method were re-confirmed by molecular methods. By comparing the phenotypic finding with the genotypic results, we identified 80 (64%) Klebsiella isolates as ESBL positive and 45 (36%) isolates as ESBL negative. These genotypic findings of the ESBL producing strains were similar to the results obtained from the combination disc method. It also confirmed that the double disc synergy test was unable to detect 47 ESBL producing klebsiella strains. Conclusion: For implementing strict infection control policy and to stop the spread of resistant strains, it is important to detect ESBL producing isolates more rapidly. A suitable antimicrobial therapy can be started timely with the early detection of resistance enzymes either phenotypically or genotypically. It has been proved that the combination disc test can be used more reliably and preferably as compared to the double disc synergy test for the detection of ESBL enzymes produced by Klebsiella strains.

scholarly journals ANTIMICROBIAL SUSCEPTIBILITY AND DETECTION METHODS FOR THE EXTENDED‑SPECTRUM β-LACTAMASES PRODUCING ENTEROBACTERIACEAE FROM CLINICAL SAMPLES

2018 ◽  
Vol 11 (5) ◽  
pp. 139
Author(s):  
Kaviyarasan G ◽  
Rajamanikandan Kcp ◽  
Sabarimuthu M ◽  
Ramya S ◽  
Arvind Prasanth D
Keyword(s):  
Infection Control ◽  
Bacterial Infections ◽  
Control Measures ◽  
Detection Methods ◽  
Clinical Samples ◽  
Healthcare Settings ◽  
Infection Control Measures ◽  
Extended Spectrum ◽  
Drug Choice ◽  
Synergy Test

Objectives: Detection of extended-spectrum β-lactamases (ESBLs) is crucial for the infection control and antibiotic choice in healthcare settings. The aim of this study is to develop a standardized, inexpensive, and simple approach that is able to detect ESBL-producing Enterobacteriaceae isolates.Methods: Isolates those were resistant to at least one of the three indicator cephalosporins (cefotaxime, cefpodoxime, and ceftazidime) were tested for ESBL production using the double disc synergy test (DDST), combined disc synergy test (CDST) test and genotypic detection of the responsible gene for the ESBL.Result: From 64 isolates, 28 were resistant to cephalosporins. In 28 isolates, 23 were positive in CDST but in the DDST 18 were showing ESBL positive. 10 were positive in both CDST and DDST.Conclusion: Resistance to cephalosporins, which are the drug choice to treat mixed bacterial infections by the Enterobacteriaceae of which disseminate rapidly being plasmid mediated. Hence, it is necessary that rapid detection of ESBL should be done and immediate infection control measures should be implemented to prevent their dissemination.

scholarly journals Detection and Antimicrobial Susceptibility Pattern of Extended Spectrum Beta Lactamases (ESBLs) Producing Gram Negative Bacteria from Different Clinical Samples

Pulse ◽  
2016 ◽  
Vol 8 (1) ◽  
pp. 15-20
Author(s):  
Suma Mita Biswas ◽  
Nikhat Ara ◽  
Nasreen Huda ◽  
Sofia Andalib ◽  
Mohammad Hefzur Rahman ◽  
...  
Keyword(s):  
Antibiotic Susceptibility ◽  
Tertiary Care ◽  
Detection Methods ◽  
Clinical Samples ◽  
Care Hospital ◽  
Clinical Diagnostic Laboratory ◽  
Diagnostic Laboratory ◽  
Gram Negative ◽  
Extended Spectrum ◽  
Antibiotic Susceptibility Test

Introduction: Extended spectrum ?-lactamases (ESBLs) are enzymes that mediate resistance to extended-spectrum (third generation) cephalosporins (e.g., ceftazidime, cefotaxime, and ceftriaxone) and monobactams (e.g., aztreonam) but do not affect cephamycins (e.g., cefoxitin and cefotetan) or carbapenems (e.g., meropenem or imipenem). Though the no. of ESBLs producing organism has been increasing day by day, the detection methods and treatment option for them are extremely limited.Aims & Objective: The present study was undertaken to investigate the rate of ESBLs production and their antibiotic susceptibility pattern.Materials & Method: A total 110 Gram negative isolates from various clinical samples from a tertiary care hospital were studied and ESBLs production was detected by double disc synergy test. Antibiotic susceptibility test was done for commonly used antibiotics.Results: Among the total isolates 66.36% (73) were ESBLs producer, and the rate of ESBLs positivity was 80.32% for E.coli (49 out of 61), 25% for Pseudomonas spp (6 out of 24), 71.42% for Klebsiella spp (10 out of 14), 80% for Enterobacter spp (4 out of 5), 100% for Acinetobacter spp (4 out of 4) and 0% for Proteus spp (0 out of 2). ESBLs producing organisms were resistant to most of the antibiotics but 100% were sensitive to imepenem.Conclusion: Screening for ESBLs production needs to be carried out routinely in every clinical diagnostic laboratory to guide clinicians in proper selection of antibiotics.Pulse Vol.8 January-December 2015 p.15-20

scholarly journals Prevalence and Antibiogram of Generic Extended-Spectrum β-Lactam-Resistant Enterobacteria in Healthy Pigs

2015 ◽  
Vol 7 (3) ◽  
pp. 272-280 ◽  
Author(s):  
Ifeoma Chinyere UGWU ◽  
Madubuike Umunna ANYANWU ◽  
Chidozie Clifford UGWU ◽  
Ogbonna Wilfred UGWUANYI
Keyword(s):  
Antibacterial Agents ◽  
Tetracycline Resistance ◽  
Diffusion Method ◽  
Phenotypic Characterization ◽  
Generic Level ◽  
Klebsiella Species ◽  
Phenotypic Resistance ◽  
E Coli ◽  
Extended Spectrum ◽  
Enugu State

This study was conducted to isolate generic extended-spectrum β-lactam (ESBL)-resistant enterobacteria from pigs reared in Enugu State Southeast, Nigeria and determine the antibacterial resistance profile of the isolates. Rectal swabs were collected from 190, randomly selected, apparently healthy pigs. Isolation of ESBL-resistant enterobacteria was done using Mac Conkey agar supplemented with 2 µg/ml of cefotaxime. Phenotypic characterization of the isolates to generic level was done following standard biochemical methods. Phenotypic resistance of the isolates to antibacterial agents was determined using the disc diffusion method. Out of 46 ESBL-resistant enterobacterial isolates, 4 (8.7%) were Escherichia coli, 11 (23.9%) were Salmonella species, while 31 (67.4%) were Klebsiella species. Resistance of the Salmonella isolates was 45.5% to ciprofloxacin, 36.4% to ofloxacin and levofloxacin, 9.1% to norfloxacin, amikacin and gentamicin, 27.3% to streptomycin, 72.7% to chloramphenicol and 90.9% to tetracycline. Resistance of the Klebsiella isolates was 93.5% to ampicillin, 12.9% to ciprofloxacin, 19.4% to ofloxacin and levofloxacin, 9.7% to norfloxacin and streptomycin, 64.5% to chloramphenicol and 38.7% to tetracycline. Resistance of the E. coli isolates was 100% to gentamicin, 75% to ampicillin and streptomycin, 50% to ciprofloxacin, norfloxacin, chloramphenicol and tetracycline, and 25% to ofloxacin, levofloxacin and amikacin. All the isolates were resistant to ceftriaxone, cefotaxime, ceftazidime, cefepime, cefpodoxime, amoxicillin/clavulanic acid and aztreonam. Resistance of the isolates to more than 3 classes of antibacterial agents tested was 54.8% for Klebsiella, 90.9% for Salmonella and 100% for E. coli, respectively. This study has shown that pigs reared in Enugu State Southeast, Nigeria, are colonized by ESBL-resistant Enterobactericeae and are potential reservoirs and disseminators of these organisms.

scholarly journals Detection of CTX-M gene in extended spectrum beta lactamase (ESBL) producing Escherichia coli and Klebsiella species of different hospitals.

2012 ◽  
Vol 4 (2) ◽  
pp. 28-31 ◽  
Author(s):  
Nawshad Muhammad Wahidur Rahman ◽  
Afzalunnessa Binte Lutfor ◽  
Sanya Tahmina Jhora ◽  
Mahmuda Yasmin ◽  
Jalaluddin Ashraful Haq
Keyword(s):  
Escherichia Coli ◽  
Tertiary Care ◽  
Beta Lactamase ◽  
M Gene ◽  
Klebsiella Species ◽  
Extended Spectrum Beta Lactamase ◽  
Extended Spectrum ◽  
Synergy Test ◽  
Tertiary Care Hospitals ◽  
Klebsiella Spp

A total of 200, non-duplicate ESBL producing strains (171 Escherichia coli and 29 Klebsiella spp.) from three tertiary care hospitals were detected using screening test & double disc synergy test. All isolates were screened for the detection of CTX-M type Extended spectrum Beta-lactamase (ESBL) using PCR. Among them 133 (66.5%) were positive for CTX-M type ESBLs which include 114 (66.66%) E.coli and 19 (65.51%) Klebsiella spp. This is the first report of identifying CTX-M gene in ESBL producing Escherichia coli and Klebsiella species of different hospitals.DOI: http://dx.doi.org/10.3329/bjmm.v4i2.10829 

scholarly journals The aadB Gene Cassette Is Associated with blaSHV Genes in Klebsiella Species Producing Extended-Spectrum β-Lactamases

2005 ◽  
Vol 49 (2) ◽  
pp. 794-797 ◽  
Author(s):  
Louisa A. Jones ◽  
Christopher J. McIver ◽  
Mi-Jurng Kim ◽  
William D. Rawlinson ◽  
Peter A. White
Keyword(s):  
Gene Cassette ◽  
Amplicon Sequencing ◽  
Klebsiella Species ◽  
Class 1 Integron ◽  
Beta Lactamases ◽  
Gene Cassettes ◽  
Extended Spectrum ◽  
Class 1 ◽  
Extended Spectrum Beta Lactamases

ABSTRACT Integrons were detected in 37 (72.5%) of 51 Klebsiella spp. producing extended-spectrum beta-lactamases by PCR with primers that targeted integrase genes and cassette regions. PCR and amplicon sequencing of the cassette regions revealed aadB and aadA2 gene cassettes that confer resistance to a range of aminoglycosides. aadB was associated with a class 1 integron on a 28-kb plasmid, pES1, that also contained bla SHV-12 and IS26.

scholarly journals COMPARISON BETWEEN DOUBLE DISK SYNERGY TEST (DDST) AND INHIBITORY POTENTIATED DISK DIFFUSION TEST (IPDDT) FOR DETECTION OF ESBL ISOLATES.

2021 ◽  
pp. 130-132
Author(s):  
Sushil Kumar Sahu ◽  
Anshu Sharma ◽  
Neha Agrawal ◽  
Subhangi Sharma
Keyword(s):  
Clavulanic Acid ◽  
Pleural Fluid ◽  
Disk Diffusion ◽  
Clinical Samples ◽  
Esbl Production ◽  
Diffusion Test ◽  
Klebsiella Species ◽  
Disk Diffusion Test ◽  
E Coli ◽  
Synergy Test

Objective: To establish effectiveness between Double Disk Synergy Test (DDST) and Inhibitory Potentiated Disk Diffusion Test (IPDDT) for detection of ESBL isolates.Material and Method: Total two hundred screened isolation of E. coli and Klebsiella species obtained from 3146 various clinical samples such as pus, urine, blood, CSF, ear discharge, pleural fluid & sputum were included in the study. ESBL production where detected by Double Disk Synergy Test (DDST) and Inhibitory Potentiated Disk Diffusion Test (IPDDT) as per CLSI guidelines.Results: 200 isolates 117(58.5 %) E.coli and 83(41.5%) Klebsiella were identified.By DDST test 92.6% & 89.1%,66.7% & 71.7%,88.8% & 71.7% showed synergy between Ceftazidime, Cefotaxime, Ceftriaxone and Amoxicillin with Clavulanic acid by Klebsiella species and E.coli respectively.By IPDDT test 27.7% & 40.1%,28.9% & 43.4% and 36.1% & 44.4% showed synergy between Ca Vs Ca/C , Ce Vs Ce/C ,Ca Vs Ca/C + Ce Vs Ce/C by Klebsiella species and E.coli respectively.The ESBL isolate detected by DDST was 73(36.5%) and by IPDDT was 82 (41%). Conclusion: Additional 4.5% (9) ESBL isolates detected by IPDDT than DDST. ESBL detection have a significant impact on several important clinical outcomes. ESBLs outbreaks require accurate and precise detection.

scholarly journals Antimicrobial Resistance Patterns in Clinical Isolates of Enterobacteriaceae from a Tertiary Care Hospital, Kathmandu, Nepal

2018 ◽  
Vol 1 (2) ◽  
pp. 74-78
Author(s):  
Ram Prasad Adhikari ◽  
Subha Shrestha ◽  
Junu Richhinbung Rai ◽  
Ritu Amatya
Keyword(s):  
Multidrug Resistance ◽  
Antimicrobial Resistance ◽  
College Teaching ◽  
Clinical Isolates ◽  
High Rate ◽  
Clinical Samples ◽  
Care Hospital ◽  
Beta Lactamases ◽  
Extended Spectrum ◽  
Extended Spectrum Beta Lactamases

Introduction: Multidrug resistance among Enterobacteriaceae is in increasing trend these days. The objective of this study was to determine the antibiogram of clinical isolates of Enterobacteriaceae with special reference to multidrug resistance and  extended spectrum beta-lactamases production.Materials and Methods: A descriptive cross sectional study was conducted over a period of six months (February -July, 2017) in the microbiology laboratory of Nepal Medical College Teaching Hospital, Kathmandu, Nepal. A total of 936 bacterial isolates of Enterobacteriaceae from clinical specimens were processed for antimicrobial susceptibility testing and screened for multidrug resistance. ESBL production was detected among potential isolates by combination disk diffusion test.Results: The rate of multidrug resistance and extended spectrum beta-lactamases production was 54.2% and 23.8% respectively. Of the total ESBL producers 92.4% were multidrug resistance. The rate of multidrug resistance and extended spectrum beta-lactamases production were higher in organisms isolated from clinical samples collected from inpatients. High rate of multidrug resistance and extended spectrum beta-lactamases production was seen in E. coli (54.4% & 27.7%), Klebsiella spp. (67.1% & 28.2%) and Citrobacter spp. (70.3% & 10.9%). The antimicrobial resistance rate was highest against ampicillin (76.7%) followed by cefixime (54. 0%), ceftazidime (51.5%), ceftriaxone (51.0%), cotrimoxazole (48.7%), ciprofloxacin (43.9%) and ofloxacin (41.1%).Conclusions:  Multidrug resistance is common among Enterobacteriaceae. These bacteria have high rate of resistance against commonly used groups of antibiotics like cephalosporins and quinolones. Continuous monitoring, surveillance of antimicrobial resistance, proper infection control and practices are important to combat with these issues.

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