scholarly journals Isolation of Metallo Beta Lactamases from Clinical Sample (MBL)

2021 ◽  
pp. 68-73
Author(s):  
Jashmi Chandraker ◽  
Ashish Saraf ◽  
Sachin Chandraker ◽  
Kanupriya Dubey
Keyword(s):  
Clinical Sample ◽  
Outer Membrane Proteins ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Beta Lactamase ◽  
Gram Negative ◽  
Beta Lactamases ◽  
Extended Spectrum Beta Lactamase ◽  
Carbapenem Resistant ◽  
Microbiological Methods

The rapid spread of acquired metallo-beta-lactamases (MBLs) among major Gram-negative pathogens is an emerging threat and a matter of particular concern worldwide. Carbapenems are among the  few useful antibiotics against gram negative bacteria particularly those  with extended spectrum beta-lactamase. Resistant to carbapenem is mediated by loss of outer membrane proteins and production of beta lactamase that is capable of hydrolyzing carbapenems.During this study period, 80 different clinical samples were collected from various hospitals of Raipur. All clinical samples were processed according to standard microbiological methods. Isolated GNRs were subjected to susceptibility testing against antibiotics by disc diffusion method according to the Clinical and Laboratory Standards Institute (CLSI) guidelines.  Carbapenem-resistant isolates were subjected to the detection of MBL production by different methods.  Out of 80 samples, 28 (35%) samples showed significant growth. MBL production was observed in 20 samples.

scholarly journals PREVALENCE OF CEPHALOSPORIN-RESISTANT GRAM-NEGATIVE BACILLI FROM CLINICAL SAMPLES

2016 ◽  
pp. 176
Author(s):  
Kavi Aniis ◽  
Rajamanikandan Kcp ◽  
Arvind Prasanth D
Keyword(s):  
Clinical Samples ◽  
Beta Lactamase ◽  
Bacterial Isolates ◽  
Beta Lactam ◽  
Gram Negative ◽  
Beta Lactamases ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Lactam Ring ◽  
Esbl Producers

<p>ABSTRACT<br />Objective: Beta-lactams are the group of antibiotics that contain a ring called as “beta-lactam ring,” which is responsible for the antibacterial activity.<br />The presence of resistance among Gram-negative organisms is due to the production of beta-lactamases enzymes that hydrolysis the beta-lactam ring<br />thereby conferring resistance to the organism. This study is undertaken to determine the prevalence of extended-spectrum beta-lactamase (ESBL)<br />producing Gram-negative organism from clinical samples.<br />Methods: A total of 112 clinical samples were taken for this study. The combined disc synergistic test (CDST) was used for the phenotypic detection<br />of ESBL producers from the clinical samples. The genotypic identification of ESBL producers was carried out by alkaline lysis method by isolation of<br />plasmid DNA.<br />Result: A total of 87 bacterial isolates were isolated and identified. Among them, Klebsiella (41%) was the predominant organism followed by<br />Escherichia coli (33%), Proteus (10%), Pseudomonas (10%), and Serratia (6%). Among the various bacterial isolates, Klebsiella showed a higher<br />percentage of resistance. The CDST showed that 8 isolates of Klebsiella, 3 isolates of E. coli, and 1 isolate of Pseudomonas were found to be ESBL<br />producers. The genotypic confirmation showed that the two bacterial isolates, namely, Klebsiella and E. coli were found to possess temoniera (TEM)<br />gene which was the 400-500 bp conferring resistance to the antibiotics.<br />Conclusion: The results of this study suggest that early detection of ESBL producing Gram-negative organism is a very important step in planning the<br />therapy of patient in Hospitals. CDST continues to be a good indicator in the detection of ESBL producers.<br />Keywords: Beta-lactamases, Gram-negative bacilli, Extended-spectrum beta-lactamase, Resistance, Combined disc synergistic test.</p><p> </p>

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.

scholarly journals Phenotypic characterization of beta-lactamases producing Gram-negative bacteria in a tertiary hospital, Nepal

2019 ◽  
Vol 7 (1) ◽  
pp. 74-81
Author(s):  
Elina Maharjan ◽  
Pooja Shakya ◽  
Balkrishna Bhattachan ◽  
Bharat Prasad Baral ◽  
Dhiraj Shrestha
Keyword(s):  
Tertiary Hospital ◽  
Diffusion Method ◽  
Phenotypic Characterization ◽  
Clinical Samples ◽  
Gram Negative Bacteria ◽  
Beta Lactamase ◽  
Gram Negative ◽  
Beta Lactamases ◽  
Disk Method ◽  
Esbl Producers

Infections caused by beta-lactamases producing Gram-negative bacteria are increasing, thus posing a challenge to the management of such infections. The surveillance data of such bacteria is limited in Nepal so this study aimed to detect the beta-lactamase producing Gram-negative bacteria in a tertiary setting. A total of 604 clinical samples, including urine, blood, sputum and body fluids, were cultured and identified by the routine standard laboratory protocols. Antibiotic susceptibility testing was done by Kirby Bauer disc diffusion method following Clinical and Laboratory Standard Institute guidelines (2014). Extended-spectrum beta-lactamases (ESBL) producers were identified by combined disk method and metallo-beta-lactamases (MBL) producers were identified by Imipenem- EDTA combined disk method. Out of 604 samples, 282 (46.7%) samples showed significant growth, of which 229 (81.2%) were Gram-negative bacteria. Of 229 Gram-negative bacteria, 200 (87.3%) were multidrug resistant, 67 (29.3%) were ESBL producers and 16 (7.0%) were MBL producers. Klebsiella pneumoniae were among higher ESBL producers and Pseudomonas aeruginosa were among higher MBL producers. The findings suggest higher antibacterial resistance among Gram-negative bacteria with the added burden of beta-lactamase production. Imipenem was effective against 125 of 229 Gram-negative bacteria tested. Thus, imipenem can be the drug of choice for empirical management. The higher multidrug resistance and higher beta-lactamases production among Gram-negative bacteria warrant the continuous monitoring, surveillance, early detection, and infection control practices of such bacteria

scholarly journals Frequency and susceptibility pattern of metallo-beta-lactamase producers in a hospital in Pakistan

2010 ◽  
Vol 4 (12) ◽  
pp. 810-813 ◽  
Author(s):  
Fatima Kaleem ◽  
Javaid Usman ◽  
Afreenish Hassan ◽  
Aslam Khan
Keyword(s):  
Tertiary Care ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Care Hospital ◽  
Gram Negative ◽  
Medical College ◽  
Strip Method ◽  
Carbapenem Resistant ◽  
Microbiological Methods ◽  
National University

Introduction: The rapid spread of acquired metallo-beta-lactamases (MBLs) among major Gram-negative pathogens is an emerging threat and a matter of particular concern worldwide. Methodology: This descriptive study was conducted between January and August 2009 in the department of Microbiology, Army Medical College, National University of Sciences and Technology, Rawalpindi, to determine the frequency and susceptibility patterns of MBL-producers among carbapenem-resistant Gram-negative rods (GNRs) from clinical isolates of a tertiary care hospital. All clinical samples were processed according to standard microbiological methods. Isolated GNRs were subjected to susceptibility testing against various antibiotics by disc diffusion method according to the Clinical and Laboratory Standards Institute (CLSI) guidelines.  Carbapenem-resistant isolates were subjected to the detection of MBL production by the E-test MBL strip method. Results: Out of 50 carbapenem resistant isolates, 39 (78%) of were confirmed to be MBL producers by the E-strip method.  Acinetobacter baumannii were the most frequent MBL producers, followed by Pseudomonas aeruginosa. A total of 19 (37%) of the MBL producers were susceptible to cefoperazone-sulbactam. Conclusion: The findings strongly suggest that there is a need to track the detection of MBL producers and that judicious use of carbapenems is necessary to prevent the further spread of these organisms.

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 First Detection of Carbapenem-Resistant Escherichia fergusonii Strains Harbouring Beta-Lactamase Genes from Clinical Samples

Pathogens ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 164 ◽  
Author(s):  
Tomilola Adesina ◽  
Obinna Nwinyi ◽  
Nandita De ◽  
Olayemi Akinnola ◽  
Emmanuel Omonigbehin
Keyword(s):  
16S Rrna ◽  
16S Rrna Sequencing ◽  
Clinical Samples ◽  
Sequencing Analysis ◽  
Beta Lactamase ◽  
Extended Spectrum Beta Lactamase ◽  
Carbapenem Resistant ◽  
Evolutionary Relatedness ◽  
Rrna Sequencing ◽  
Physiological Tests

Recently discovered extraintestinal Escherichia fergusonii obtained from non-clinical samples has exhibited the potential for acquiring multiple beta-lactamase genes, just like many extraintestinal Escherichia coli strains. Albeit, they are often omitted or classified as E. coli. This study aimed to, therefore, identify carbapenem-resistant extended-spectrum beta-lactamase (ESBL) producing E. fergusonii isolates from clinical samples, determine their evolutionary relatedness using 16S rRNA sequencing analysis and screen for beta-lactamase genes. A total of 135 septic wound samples were obtained from patients on referral at a General Hospital in Lagos, Nigeria. For the phenotypic identification of isolates from culture-positive samples, morphological, and physiological tests were carried out. Identities of the isolates harbouring beta-lactamase genes were assigned to their genus strains using the 16S rRNA sequencing. The Kirby Bauer disc diffusion technique and double-disc synergy test were used to screen isolates for multidrug resistance and ESBL production. Carbapenem-resistant ESBL producing isolates were screened for beta-lactamase genes in a polymerase chain reaction. Three E. fergusonii isolates (CR11, CR35 and CR49) were obtained during this study. E. fergusonii strains were motile, non-lactose and non-sorbitol fermenting but positive for cellobiose and adonitol fermentation. The I6S rRNA assigned the phenotypically identified isolates to E. fergusonii species. All three isolates were multidrug-resistant, carbapenem-resistant and ESBL producers. Isolates CR11 and CR35 harboured cefotaximase (CTX-M) and temoniera (TEM) beta-lactamase genes while CR49 harboured sulfhydryl variable (SHV) beta-lactamase gene. We herein report the detection of multiple beta-lactamase genes in carbapenem-resistant ESBL producing E. fergusonii from clinical samples.

scholarly journals Bad Bugs and No Drugs: Activity of Colistin as Waging War against Emerging Metallo-β-Lactamases Producing Pathogens

2018 ◽  
Vol 24 (1) ◽  
pp. 625-631 ◽  
Author(s):  
Sahar Naz ◽  
Farhan Rasheed ◽  
Muhammad Saeed ◽  
Shagufta Iram ◽  
Ambereen Anwar Imran
Keyword(s):  
Cross Sectional Study ◽  
Resistant Isolate ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Klebsiella Species ◽  
Cross Sectional ◽  
Acinetobacter Species ◽  
Gram Negative ◽  
Negative Results ◽  
Carbapenem Resistant

Inter-hospital and intra-hospital dissemination of metallo-β-lactamase (MβL) producing strains possess significant therapeutic challenges. Objective: This study was carried out to evaluate the efficacy of Colistin against MβL producers. Material and Methods: This cross-sectional study was conducted in Microbiology Laboratory, Allama Iqbal Medical College, Lahore, Pakistan from 1stJuly 2016 to 25th February 2017. A total of 12126 clinical samples were collected from patients presenting to Jinnah Hospital, Lahore. Every sample was processed for bacterial culture. Bacterial identification was performed according to standard guidelines. Every gram-negative isolate was further processed for antimicrobial susceptibility testing by modified Kirby Baur disc diffusion method. Zone sizes were interpreted according to CLSI 2016 guidelines. Next day every carbapenem-resistant isolate were further processed for MβL detection by EDTA method, zone size of Carbapenem disc only and Carbapenem disc impregnated with EDTA was compared ( >7 mm increase MβL positive, 0-5 mm increase MβL-negative). Results: Out of total 12126 samples, 35.9% (n=4361) were culture positive and only 40.5% (n=1770) were Gram negative rods. Of these 9.6% (n=170) were Carbapenem-resistant isolates with 47% (n=80) MβL producers. Briefly 51.7% (n=30) Acinetobacter species were MβL positive, Pseudomonas species 38.5% (n=22), Escherichia coli 69.5% (n=16), Klebsiella species 37.0% (n=10), Proteus 66.6% (n=2) and 0% Citrobacter sppwere MβL positive. 32.5% MβL positive isolates were from ICU, 21.2% were from OPD, 12.5%were from Surgical Units, 12.5% were from Medical Unit, 17.5% were from Orthopedic Unit, and 3.7% were from Pulmonology ward. Almost 100% resistant was observed in MβL positive isolates for Imipenem,Piperacillin+ Tazobactum, Ceftriaxone, Co-amoxyclav, Cefoperazone+Sulbactam, Ciprofloxacin, and Amikacin, Doxycycline, and Gentamicin showed 91.2%, 94.0%, and 97.5% resistant rate respectively. No resistance was observed against Colistin. Conclusion: MβL producing Gram negative rods are rising in clinical setups. They are becoming a nightmare for clinicians to treat such infections. Colistin remains the only choice of drug for MβL positive and Negative isolates with 0% resistant rate except for Proteus species, to which it is intrinsically resistant.

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