scholarly journals Identification and Antibiotic Susceptibility Testing of ESBL producing Klebsiella strains by Phenotypic methods

2018 ◽  
Vol 9 (1) ◽  
pp. 8-13
Author(s):  
Malik Taqdees ◽  
Asma Naim ◽  
Asma Saeed
Keyword(s):  
Diffusion Method ◽  
Multi Drug Resistance ◽  
Beta Lactamase ◽  
Disk Diffusion Method ◽  
Klebsiella Species ◽  
Gram Negative ◽  
Extended Spectrum Beta Lactamase ◽  
Extended Spectrum ◽  
Klebsiella Spp ◽  
Esbl Producers

Multi drug resistance has now become a worldwide therapeutic challenge due to the widespread use of broad spectrum antibiotics. Klebsiella species have significant importance in clinical field as they cause various infections in human and are considered as potential pathogens that express antibiotic resistance through their strong enzymatic activity. Extended spectrum beta lactamases (ESBLs) are plasmid mediated enzymes produced mostly because of mutation and few other factors.  These enzymes confer resistance against various β-lactam drugs including cephalosporins and monobactams. Among the genus Klebsiella, ESBLs are highly prevalent in K. pneumoniae followed by K. oxytoca. This study was conducted in Pakistan to assess the distribution of ESBL producers among Klebsiella spp., an important member of the family Enterobacteriaceae. From January 2010 to January 2012, a total of 236 gram-negative isolates were collected from different renowned microbiological laboratories. Out of the 236 gram-negative isolates, 125 were found as Klebsiella spp. by using standard microbiological techniques. Antimicrobial susceptibility profiling of these strains was performed by using Kirby Bauer disk diffusion method. Phenotypic detection of the production of extended spectrum beta lactamase enzyme was performed using double disc synergy method and combination disc method. It has been identified that Klebsiella strains are highly resistant against Amoxicillin, Tetracycline, Nalidixic Acid, Cephradine, Gentamicin, co-amoxyclav with the percentage of 100%, 86%, 86%, 82%, 82% and 80% respectively. The most effective antibiotics for Klebsiella spp. were found to be Amikacin, Meropenem and Piperacillin-tazobactam, with highest sensitivities of 96%, 94% and 91%. Phenotypic detection of Extended spectrum beta lactamase production by double disc synergy test was able to identify 28% ESBL producers among Klebsiella isolates whereas 64% were detected by combination disc test.

scholarly journals Evaluation of extended spectrum beta lactamase enzymes prevalence in clinical isolates of Escherichia coli

2011 ◽  
Vol 2 (1) ◽  
pp. 8
Author(s):  
Ronak Bakhtiari ◽  
Jalil Fallah Mehrabadi ◽  
Hedroosha Molla Agamirzaei ◽  
Ailar Sabbaghi ◽  
Mohammad Mehdi Soltan Dallal
Keyword(s):  
Escherichia Coli ◽  
Disk Diffusion ◽  
Confirmatory Test ◽  
Diffusion Method ◽  
Multi Drug Resistance ◽  
Beta Lactamase ◽  
Disk Diffusion Method ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Extended Spectrum

Resistance to b-lactam antibiotics by gramnegative bacteria, especially <em>Escherichia coli (E. coli)</em>, is a major public health issue worldwide. The predominant resistance mechanism in gram negative bacteria particularly <em>E. coli </em>is via the production of extended spectrum beta lactamase (ESBLs) enzymes. In recent years, the prevalence of b-lactamase producing organisms is increased and identification of these isolates by using disk diffusion method and no-one else is not satisfactory. So, this investigation focused on evaluating the prevalence of ESBL enzymes by disk diffusion method and confirmatory test (Combined Disk). Five hundred clinical samples were collected and 200 <em>E. coli </em>isolates were detected by standard biochemical tests. To performing initial screening of ESBLs was used from Disk diffusion method on <em>E. coli </em>isolates. A confirmation test (Combined Disk method) was performed on isolates of resistant to cephalosporin's indicators. Up to 70% isolates exhibited the Multi Drug Resistance phenotype. In Disk diffusion method, 128(64%) <em>E. coli </em>isolates which resistant to ceftazidime and cefotaxime while in Combined Disk, among 128 screened isolates, 115 (89.8%) isolates were detected as ESBLs producers. This survey indicate beta lactamase enzymes are playing a significant role in antibiotic resistance and correct detection of them in phenotypic test by using disk diffusion and combined Disk is essential for accurate recognition of ESBLs.

Prevalence and Antibiogram of Extended-Spectrum Beta-Lactamase Producing Gram-negative Bacteria Isolated from Septicemic Neonates in a Tertiary Care Hospital

KYAMC Journal ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 171-175
Author(s):  
Tania Rahman ◽  
Momtaz Begum ◽  
Sharmeen Sultana ◽  
SM Shamsuzzaman
Keyword(s):  
Antimicrobial Susceptibility ◽  
Diffusion Method ◽  
Gram Negative Bacteria ◽  
Care Hospital ◽  
Beta Lactamase ◽  
Blood Samples ◽  
Gram Negative ◽  
Extended Spectrum Beta Lactamase ◽  
Extended Spectrum ◽  
Esbl Producers

Background: In recent years, Extended-spectrum beta-lactamase (ESBL) producing microorganisms have complicated treatment of infections due to resistance of ESBL producing strains to a wide range of antimicrobials. Objective: Target of this study was to determine the prevalence of ESBL producing gramnegative bacteria in neonatal sepsis cases and to reveal the antimicrobial susceptibility pattern of those isolated ESBL producers. Materials and Methods: This cross sectional study was carried out in Dhaka Medical College Hospital (DMCH) over a period of 12 months from January to December in 2016. Following isolation and identification of gram-negative bacteria from blood samples of suspected septicemic neonates, antimicrobial susceptibility test was performed by Kirby Bauer disk-diffusion method and ESBL producers were detected by Double Disk Synergy (DDS) test. Results: Among 52 Gram-negative bacteria isolated from 106 blood samples, 34.61% ESBL producers were detected and Enterobacter spp. (45%) was predominant followed by Klebsiella pneumoniae (33.33%). None of the ESBL producers was resistant to colistin and tigecycline. All ESBL producing Acinetobacter baumannii, 77.78% and 66.67% of ESBL producing Enterobacter spp and Klebsiella spp. respectively showed resistance to meropenem. All ESBL producers were resistant to piperacillintazobactam. Conclusion: Appropriate measures should be taken to prevent the spread of ESBL producing strains by combining strategies for infection prevention, control and rational use of antibiotics. KYAMC Journal Vol. 11, No.-4, January 2021, Page 171-175

scholarly journals Phenotypic and Genotypic Study of Klebsiella species with Reference to Extended Spectrum Beta-Lactamase

2021 ◽  
Author(s):  
Rakesh Prasad Sah ◽  
Rakesh Kumar Mukhia ◽  
AD Urhekar ◽  
Kshitija Rane
Keyword(s):  
Drug Resistance ◽  
Multi Drug Resistance ◽  
Pcr Analysis ◽  
Beta Lactamase ◽  
Klebsiella Species ◽  
Single Strain ◽  
Antimicrobial Drug Resistance ◽  
Extended Spectrum Beta Lactamase ◽  
Extended Spectrum ◽  
Esbl Producers

Introduction: Klebsiella species is an important nosocomial pathogen with the emergence of Multi Drug Resistance (MDR). MDR in Klebsiella species is increasing worldwide with the production Extended Spectrum Beta-Lactamase (ESBL). The emergence of ESBL is a critical concern in Klebsiella species due to resistance to ceftazidime and other cephalosporins which compromise the efficacy of life saving antibiotics against these infections. Aim: To know the factors responsible for antimicrobial drug resistance in Klebsiella species with respect to ESBL and their responsible genes. Materials and Methods: A prospective and experimental study was carried out over a period of three years (August 2013 to July 2016). Total 200 isolates of Klebsiella species were screened for cefotaxime and ceftazidime. The resistant strains (cefotaxime/ ceftazidime) were subjected to ESBL agar, Phenotypic Confirmatory Disc Diffusion Test (PCDDT) and Modified Three Dimensional Test (M3DT). Genetic analysis by Polymerase Chain Reaction (PCR) was done for the detection of beta-lactamase (bla) genes i.e., blaTEM, blaSHV & blaCTX-M in 58 isolates of Klebsiella species. The data was presented using frequency and percentage. The proportion was compared using Z-test for the proportions. Results: Out of 200 isolates, 135 (67.5%) were found resistant to cefotaxime and 125 (62.5%) were resistant to ceftazidime. Among which 110 (55%), 75 (37.5%) and 95 (47.5%) Klebsiella species were found positive for production of ESBL by ESBL agar, PCDDT and M3DT respectively. PCR analysis in 48 isolates were positive by PCDDT/M3DT or both were also positive for beta- lactamase genes i.e., 43 (89.58%) blaTEM; 44 (91.67%) blaSHV and 48 (100%) blaCTX-M. Ten negative isolates either by PCDDT/M3DT or both were also negative by PCR. Co-existence of (blaTEM+blaSHV +blaCTX-M), (blaTEM+blaSHV), (blaTEM+blaCTX-M) and (blaSHV+blaCTX-M) were found 81.25%, 0%, 8.33% and 10.42%, respectively. Conclusion: The M3DT is the best phenotypic method for the confirmation of ESBL producer in Klebsiella species which is not included by CLSI while inclusion with PCDDT enhances the detection of ESBL producers. Co-existence of all three genes (blaTEM, blaSHV and blaCTX-M) in a single strain is a serious concern for us. So it is important to include M3DT and PCDDT in routine basis for the detection and management of ESBL producers which will help clinician to prescribe proper antibiotics.

scholarly journals Detection of Extended Spectrum Beta-lactamase (ESBL) Producing Gram Negative Bacteria from Clinical Specimens of Sir Salimullah Medical College and Mitford Hospital.

2017 ◽  
Vol 10 (1) ◽  
pp. 8-12
Author(s):  
Shikha Paul ◽  
Sanya Tahmina Jhora ◽  
Prashanta Prasun Dey ◽  
Bilkis Ara Begum
Keyword(s):  
Tertiary Care ◽  
Gram Negative Bacteria ◽  
Care Hospital ◽  
Beta Lactamase ◽  
Gram Negative ◽  
Clinical Specimens ◽  
Extended Spectrum Beta Lactamase ◽  
Medical College ◽  
Extended Spectrum ◽  
Esbl Producers

Detection of Extended spectrum beta lactamase (ESBL) enzyme producing bacteria in hospital settings is vital as ESBL genes are transmissible. This study was carried out to determine the distribution of ESBL producing gram negative isolates at a tertiary care hospital in Dhaka city which deals with the patients hailing from relatively low socioeconomic status.Onehundred and twenty four gram negative bacteria isolated from different clinical specimens from outpatient and inpatient departments of Sir Salimullah Medical College and Mitford Hospital (SSMC & MH) were tested for ESBL by E test ESBL method in the department of microbiology of Sir Salimullah medical college (SSMC) from March 2013 to August 2013.Out of 124 gram negative bacteria 69 (55.65%) were positive for ESBL. Among the ESBL producers, Esch.coli was the highest (46.38%) which was followed by Serratia spp (11.59%), Enterobacter spp (10.14%), Proteus spp, (8.70%), Acinetobacter spp.(7.24%) and Klebsiella spp.(5.79%). Out of 32 Esch.coli isolated from outpatient department, 10 (31.25%) were positive for ESBL. On the other hand out of 27 Esch. coli isolated from inpatient department, 22 (81.48%) were positive for ESBL. The difference was statistically significant (p<0.001).So the present study reveals that the distribution of ESBL producers is more among the hospitalized patients than the patients of the community.Bangladesh J Med Microbiol 2016; 10 (1): 8-12

scholarly journals Presence of different beta-lactamase classes among clinical isolates of Pseudomonas aeruginosa expressing AmpC beta-lactamase enzyme

2010 ◽  
Vol 4 (04) ◽  
pp. 239-242 ◽  
Author(s):  
Supriya Upadhyay ◽  
Malay Ranjan Sen ◽  
Amitabha Bhattacharjee
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Treatment Options ◽  
Clinical Isolates ◽  
Diffusion Method ◽  
Beta Lactamase ◽  
Beta Lactam ◽  
Disk Diffusion Method ◽  
Extended Spectrum Beta Lactamase ◽  
Extended Spectrum ◽  
Enzyme Detection

Introduction: Infections caused by Pseudomonas aeruginosa are difficult to treat as the majority of isolates exhibit varying degrees of beta-lactamase mediated resistance to most of the beta-lactam antibiotics. It is also not unusual to find a single isolate that expresses multiple β-lactamase enzymes, further complicating the treatment options. Thus the present study was designed to investigate the coexistence of different beta-lactamase enzymes in clinical isolates of P. aeruginosa. Methodology: A total of 202 clinical isolates of P. aeruginosa were tested for the presence of AmpC beta-lactamase, extended spectrum beta-lactamase (ESBL) and metallo beta-lactamase (MBL) enzyme. Detection of AmpC beta-lactamase was performed by disk antagonism test and a modified three-dimensional method, whereas detection of ESBL was done by the combined disk diffusion method per Clinical and Laboratory Standards Institute (CLSI) guidelines and MBL were detected by the Imipenem EDTA disk potentiation test. Results: A total of 120 (59.4%) isolates were confirmed to be positive for AmpC beta-lactamase. Among them, 14 strains (7%) were inducible AmpC producers. Co-production of AmpC along with extended spectrum beta-lactamase and metallo beta-lactamase was reported in 3.3% and 46.6% isolates respectively. Conclusion: The study emphasizes the high prevalence of multidrug resistant P. aeruginosa producing beta-lactamase enzymes of diverse mechanisms. Thus proper antibiotic policy and measures to restrict the indiscriminative use of cephalosporins and carbapenems should be taken to minimize the emergence of this multiple beta-lactamase producing pathogens.

scholarly journals Extended Spectrum Beta-Lactamase Production in Uropathogens Isolated from Hospitalized Patients with Chronic Pyelonephritis

2015 ◽  
Vol 8 (1) ◽  
pp. 71-75 ◽  
Author(s):  
Olga I Chub ◽  
Aleksandr V Bilchenko ◽  
Igor Khalin
Keyword(s):  
Cross Sectional Study ◽  
Hospitalized Patients ◽  
Chronic Pyelonephritis ◽  
Diffusion Method ◽  
Beta Lactamase ◽  
Disk Diffusion Method ◽  
Cross Sectional ◽  
Extended Spectrum Beta Lactamase ◽  
Extended Spectrum ◽  
Tract Infections

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.

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 ESBL Producers in Gram Negative Isolates from Clinical Samples

2020 ◽  
Vol 14 (3) ◽  
pp. 2027-2032
Author(s):  
Mita D. Wadekar ◽  
J.V. Sathish ◽  
C. Pooja ◽  
S. Jayashree
Keyword(s):  
Treatment Options ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Beta Lactamase ◽  
Beta Lactam ◽  
Gram Negative ◽  
Extended Spectrum Beta Lactamase ◽  
Beta Lactam Antibiotics ◽  
Esbl Producers

Resistance to beta lactam antibiotics is the most common cause for beta-lactamase production. Increasing number of extended spectrum beta-lactamase (ESBL) producers has reduced the treatment options which resulted in emergence of multidrug resistant strains, treatment failure and hence increased mortality. To detect phenotypically, ESBL producers in Gram negative isolates from different samples and to know their susceptibility pattern. A retrospective study of Gram negative isolates was conducted. Total of 521 isolates were isolated from various samples. They were processed and identified by standard procedures. The antibiotic susceptibility testing was performed by Kirby- Bauer disc diffusion method using CLSI guidelines. ESBL was detected by combination disk test. A total of 521 Gram negative isolates were isolated which included E. coli, Klebsiella pneumoniae, Citrobacter spp., Enterobacter spp., Proteus spp. and Acinetobacter spp. Pseudomonas aeruginosa. Of 521 isolates tested, ESBL was detected in 329 (63.1%) isolates. These isolates showed maximum susceptibility to piperacillin- tazobactam (86%) followed by imipenem (78.4%), amikacin (63.5%), cotrimoxazole (54.4%), ciprofloxacin (51%), amoxi-clav (44.9%), cefepime (44.1%), gentamicin (38.9%), cefoxitin (34.9%) and ampicillin (19.1%). ESBL producers which are resistant to beta lactam antibiotics have become a major problem. Detection of these beta-lactamase enzymes by simple disk method and its reporting will help clinicians in prescribing proper antibiotics.

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