scholarly journals Evaluation of eazyplex® SuperBug CRE Test for Beta-Lactamase Genes Detection in Klebsiella spp. and P. aeruginosa Strains

2019 ◽  
Vol 77 (1) ◽  
pp. 99-103 ◽  
Author(s):  
Alicja Sękowska ◽  
Tomasz Bogiel ◽  
Eugenia Gospodarek-Komkowska
Keyword(s):  
Drug Resistance ◽  
Resistance Mechanisms ◽  
Multi Drug Resistance ◽  
Lamp Method ◽  
Beta Lactamase ◽  
Beta Lactam ◽  
Gram Negative ◽  
Beta Lactamases ◽  
Genes Encoding ◽  
Klebsiella Spp

AbstractThe multi-drug resistance of Gram-negative rods is one of the most important issues of present medicine. In recent years, more and more strains resistant to the majority or to all possible therapeutic options have been isolated—especially Klebsiella spp. and Pseudomonas spp. representatives. It is very important to detect strains with these phenotypes as quickly and reliably as possible. The aim of the study was to evaluate the usefulness of eazyplex® SuperBug CRE test (Amplex Diagnostics) for the detection of the most important beta-lactam resistance genes. eazyplex® SuperBug CRE test is based on the loop-mediated isothermal amplification (LAMP) method, and detects genes for the following beta-lactamases: KPC, NDM-1, VIM, OXA-48, CTX-M1, CTX-M9 and OXA-181. The study involved 87 strains. For all of the positive strains in the LAMP method, additional PCR were performed to increase the spectrum of ESBLs detected by the genes encoding for enzymes belonging to TEM and SHV families. The results obtained by the tested method and standard PCR were consistent for all Klebsiella spp. strains. The discrepancy between the evaluated test and PCR results was observed for one P. aeruginosa strain. The eazyplex® SuperBug CRE test can be used for quick detection of the most important beta-lactam resistance mechanisms amongst Gram-negative rods.

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 Prevalence of Extended Spectrum Beta Lactamases (ESBL) and Metallo Beta Lactamases (MBL) Mediated Resistance in Gram Negative Bacterial Pathogens

2018 ◽  
Vol 4 ◽  
pp. 49-54 ◽  
Author(s):  
Pramila Pathak ◽  
Nandalal Jaishi ◽  
Binod Kumar Yadav ◽  
Pradeep Kumar Shah
Keyword(s):  
Drug Resistance ◽  
Hospital Setting ◽  
Multi Drug Resistance ◽  
Microbiological Analysis ◽  
Bacterial Isolates ◽  
Biochemical Tests ◽  
Gram Negative ◽  
Beta Lactamases ◽  
Clinical Specimens ◽  
Extended Spectrum

Objectives: This study was conducted to determine the prevalence of multi-drug resistance (MDR) along with Extended Spectrum β-lactamase (ESBL) and Metallo β-lactamase (MBL) producing gram negative bacterial isolates among the patients attending Shahid Gangalal National Heart Centre, Kathmandu, Nepal.Methods: This cross-sectional study was carried out from June to December; 2016. Altogether 977 clinical specimens were processed for analysis of bacteriological profile and the isolates were identified by culture, morphological and biochemical tests. Antibiotic susceptibility testing of the isolates was performed by Kirby Bauer disc diffusion methods following Clinical and Laboratories Standard Institute guideline and the isolates were tested for ESBL and MBL by combined disk method.Results: out of 977 clinical specimens, 254 (25.99%) were found to be gram negative bacterial isolates, among them Klebsiella pneumoniae 83 (32.67%) was the most predominant organism followed by E. coli 51 (20.07%), Pseudomonas aeruginosa 36 (14.17%), K. oxytoca 32 (12.59%), Proteus mirabilis 13 (5.11%) and P. vulgaris 13 (5.11%), Acinetobacter spp. 11 (4.33%), Citrobacter spp. 10 (3.93%) and Enterobacter spp. 5 (1.96%) respectively. 83 (32.67%) isolates were found to be MDR, 38(14.96%) were positive for ESBL while 19 (7.48%) were MBL producer.Conclusion: The determent drug resistance among ESBL and MBL producers, reflect the extensive use of antibiotics possessing difficulties in therapeutic potions in hospital setting which might be overcome by proper microbiological analysis of pathogenic isolates and judicious use of antibiotics for emergence of resistance strains.

scholarly journals In Vitro Activity of the Ultra-Broad-Spectrum Beta-lactamase Inhibitor QPX7728 in Combination with Multiple Beta-Lactam Antibiotics against Pseudomonas aeruginosa

2021 ◽  
Author(s):  
Olga Lomovskaya ◽  
Debora Rubio-Aparicio ◽  
Kirk Nelson ◽  
Dongxu Sun ◽  
Ruslan Tsivkovski ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Resistance Mechanisms ◽  
Clinical Isolates ◽  
Beta Lactamase ◽  
In Vitro Activity ◽  
Beta Lactam ◽  
Beta Lactams ◽  
Beta Lactamases ◽  
Lactamase Inhibitor

QPX7728 is an ultra-broad-spectrum beta-lactamase inhibitor with potent inhibition of key serine and metallo beta-lactamases. QPX7728 enhances the potency of multiple beta-lactams in beta-lactamase producing Enterobacterales and Acinetobacter spp. In this study we evaluated the in vitro activity of QPX7728 (8 μg/ml) combined with multiple beta-lactams against clinical isolates of Pseudomonas aeruginosa with varying beta-lactam resistance mechanisms. Seven-hundred-ninety clinical isolates were included in this study; 500 isolates, termed a “representative panel”, were selected to be representative the MIC distribution of meropenem (MEM), ceftazidime-avibactam (CAZ-AVI), and ceftolozane-tazobactam (TOL-TAZ) resistance for clinical isolates according to 2017 SENTRY surveillance data (representative panel). An additional 290 selected isolates (“challenge panel”), that were either non-susceptible to MEM or were resistant to TOL-TAZ or CAZ-AVI were also tested; 61 strains carried metallo beta-lactamases (MBLs), 211 strains were defective in the carbapenem porin OprD and 185 strains had the MexAB-OprM efflux pump overproduced based on a phenotypic test. Against the representative panel, susceptibility for all QPX7728/beta-lactam combinations was >90%. For the challenge panel, QPX-ceftolozane (TOL) was the most active combination (78.6% susceptible) followed by equipotent QPX-piperacillin (PIP) and QPX-cefepime (FEP), restoring susceptibility in 70.3% of strains (CLSI breakpoints for the beta-lactam compound alone). For MBL-negative strains, QPX-TOL and QPX-FEP restored the MIC values to susceptibility rates in ∼90% and ∼80% of strains, respectively, vs 68-70% for QPX-MEM and QPX-PIP and 63-65% for TOL-TAZ and CAZ-AVI. For MBL-positive strains, QPX-PIP restored the MIC to susceptibility values for ∼70% of strains vs 2-40% for other combinations. Increased efflux and impaired OprD had varying effect on QPX7728 combination depending on the partner beta-lactam tested. QPX7728 enhanced the potency of multiple beta-lactams against P. aeruginosa, with varying results according to the beta-lactamase production and other intrinsic resistance mechanisms.

The Ultra-Broad-Spectrum Beta-lactamase Inhibitor QPX7728 Restores the Potency of Multiple Oral Beta-lactam Antibiotics against Beta-lactamase Producing Strains of Resistant Enterobacterales

2021 ◽  
Author(s):  
Olga Lomovskaya ◽  
Debora Rubio-Aparicio ◽  
Ruslan Tsivkovski ◽  
Jeff Loutit ◽  
Michael Dudley
Keyword(s):  
Broad Spectrum ◽  
Resistance Mechanisms ◽  
Beta Lactamase ◽  
Beta Lactam ◽  
Beta Lactams ◽  
Intrinsic Resistance ◽  
Beta Lactamases ◽  
Beta Lactam Antibiotics ◽  
The Impact ◽  
Lactamase Inhibitor

QPX7728 is a cyclic boronate ultra-broad-spectrum beta-lactamase inhibitor, with potent activity against both serine and metallo beta-lactamases. QPX7728 can be delivered systemically by the IV or oral route of administration. Oral β-lactam antibiotics alone or in combination with QPX7728 were evaluated for 1) sensitivity to hydrolysis by various common beta-lactamases and inhibition of hydrolysis by QPX7728; 2) the impact of non-beta-lactamase-mediated resistance mechanisms on potency of beta-lactams; and 3) in vitro activity against a panel of clinical strains producing diverse beta-lactamases. The carbapenem tebipenem had stability for many serine beta-lactamases from all molecular classes followed by cephalosporin ceftibuten. Addition of QPX7728 to tebipenem, ceftibuten and mecillinam completely reversed beta-lactamase-mediated resistance in cloned beta-lactamases from serine and metallo enzyme classes; the degree of potentiation of other beta-lactams varied according to the beta-lactamase produced. Tebipenem, ceftibuten and cefixime had the lowest MICs against laboratory strains with various combinations of beta-lactamases and the intrinsic drug-resistance mechanisms of porin and efflux mutations. There was a high degree of correlation between potency of various combinations against cloned beta-lactamases and efflux/porin mutants and the activity against clinical isolates, showing the importance of both inhibition of beta-lactamase along with minimal impact of general intrinsic resistance mechanisms affecting the beta-lactam. Tebipenem and ceftibuten appeared to be the best beta-lactam antibiotics when combined with QPX7728 for activity against Enterobacterales that produce serine or metallo beta-lactamases.

scholarly journals Mechanisms of Resistance in Gram-Negative Urinary Pathogens: From Country-Specific Molecular Insights to Global Clinical Relevance

Diagnostics ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 800
Author(s):  
Branka Bedenić ◽  
Tomislav Meštrović
Keyword(s):  
Clinical Importance ◽  
Resistance Mechanisms ◽  
Bacterial Strains ◽  
Mechanisms Of Resistance ◽  
Gram Negative ◽  
Beta Lactamases ◽  
Resistance Determinants ◽  
Genes Encoding ◽  
Extended Spectrum Beta Lactamases ◽  
Tract Infections

Urinary tract infections (UTIs) are the most frequent hospital infections and among the most commonly observed community acquired infections. Alongside their clinical importance, they are notorious because the pathogens that cause them are prone to acquiring various resistance determinants, including extended-spectrum beta-lactamases (ESBL); plasmid-encoded AmpC β-lactamases (p-AmpC); carbapenemases belonging to class A, B, and D; qnr genes encoding reduced susceptibility to fluoroquinolones; as well as genes encoding enzymes that hydrolyse aminoglycosides. In Escherichia coli and Klebsiella pneumoniae, the dominant resistance mechanisms are ESBLs belonging to the CTX-M, TEM, and SHV families; p-AmpC; and (more recently) carbapenemases belonging to classes A, B, and D. Urinary Pseudomonas aeruginosa isolates harbour metallo-beta-lactamases (MBLs) and ESBLs belonging to PER and GES families, while carbapenemases of class D are found in urinary Acinetobacter baumannii isolates. The identification of resistance mechanisms in routine diagnostic practice is primarily based on phenotypic tests for the detection of beta-lactamases, such as the double-disk synergy test or Hodge test, while polymerase chain reaction (PCR) for the detection of resistance genes is mostly pursued in reference laboratories for research purposes. As the emergence of drug-resistant bacterial strains poses serious challenges in the management of UTIs, this review aimed to appraise mechanisms of resistance in relevant Gram-negative urinary pathogens, to provide a detailed map of resistance determinants in Croatia and the world, and to discuss the implications of these resistance traits on diagnostic approaches. We summarized a sundry of different resistance mechanisms among urinary isolates and showed how their prevalence highly depends on the local epidemiological context, highlighting the need for tailored interventions in the field of antimicrobial stewardship.

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 In Vitro Activity of the Ultrabroad-Spectrum-Beta-Lactamase Inhibitor QPX7728 against Carbapenem-Resistant Enterobacterales with Varying Intrinsic and Acquired Resistance Mechanisms

2020 ◽  
Vol 64 (8) ◽  
Author(s):  
Kirk Nelson ◽  
Debora Rubio-Aparicio ◽  
Dongxu Sun ◽  
Michael Dudley ◽  
Olga Lomovskaya
Keyword(s):  
Acquired Resistance ◽  
Fold Increase ◽  
Resistance Mechanisms ◽  
Multiple Resistance ◽  
Beta Lactamase ◽  
Beta Lactam ◽  
Beta Lactamases ◽  
Carbapenem Resistant ◽  
Lactamase Inhibitor

ABSTRACT QPX7728 is an investigational ultrabroad-spectrum-beta-lactamase inhibitor (BLI) with potent inhibition of key serine and metallo-beta-lactamases. QPX7728 enhances the potency of many beta-lactams, including carbapenems, in isogenic strains of Gram-negative bacteria producing various beta-lactamases. The potency of meropenem alone and in combination with QPX7728 (tested at fixed concentrations of 1 to 16 μg/ml) was tested against 598 clinical isolates of carbapenem-resistant Enterobacterales (CRE). The panel included 363 strains producing serine carbapenemases, 224 strains producing metallo-beta-lactamases (151 NDM, 53 VIM, and 20 IMP), and 50 strains that did not carry any known carbapenemases but were resistant to meropenem (MIC ≥ 4 μg/ml). The panel was also enriched in strains that had various defects in the major porins OmpK35/OmpF and OmpK36/OmpC. Increasing concentrations of QPX7728 restored the potency of meropenem against CRE, with the meropenem MIC90 decreasing from >64 μg/ml to 0.5 μg/ml for QPX7728 (8 μg/ml). QPX7728 significantly increased the potency of meropenem against CRE with multiple resistance mechanisms; the reduction in the meropenem MIC90 with QPX7728 (8 μg/ml) ranged from 32- to >256-fold. Compared with other beta-lactamase inhibitor combinations, meropenem-vaborbactam, ceftazidime-avibactam, and imipenem-relebactam, meropenem with QPX7728 was the most potent beta-lactam–BLI combination tested against all groups of CRE with multiple resistance mechanisms. Defects in OmpK36 in KPC-producing strains markedly decreased the potency of meropenem with vaborbactam (128-fold increase in the MIC90), whereas only an 8- to 16-fold change was observed with QPX7728 plus meropenem. More than 90% of various CRE subsets (including those with reduced permeability) were susceptible to ≤8 μg/ml of meropenem with QPX7728 at 8 μg/ml or lower. The combination of QPX7728 with meropenem against CRE has an attractive microbiological profile in CRE with multiple resistance mechanisms.

Sulbactam/Ampicillin

1988 ◽  
Vol 9 (7) ◽  
pp. 323-327
Author(s):  
Francine R. Salamone
Keyword(s):  
Enzyme Inhibitor ◽  
The United States ◽  
Beta Lactamase ◽  
Beta Lactam ◽  
Gram Negative ◽  
Beta Lactamases ◽  
Beta Lactam Antibiotics ◽  
Development Of Resistance ◽  
Lactam Ring ◽  
Lactamase Inhibitor

Sulbactam/ampicillin was recently marketed for use in several infections caused by beta-lactamase-producing organisms. Sulbactam is the second beta-lactamase inhibitor to become available in the United States. Interest in inhibition of beta-lactamases arose in the late 1960s when a combination consisting of an antibacterial agent and an enzyme inhibitor was found effective in the treatment of certain resistant gram-negative infections. It is now well accepted that the addition of a beta-lactamase inhibitor to a beta-lactam antibiotic may expand its usefulness in a variety of infections.The penicillin derivatives, known as beta-lactam antibiotics, possess a four-membered ring (beta-lactam ring) fused to a second ring (Figure). It is the beta-lactam ring that is essential for the inhibition of bacterial cell wall synthesis and subsequent bactericidal activity of these agents. The development of resistance to beta-lactam antibiotics may occur by a number of mechanisms, although the most important is bacterial production of enzymes (beta-lactamases) that are capable of beta-lactam ring hydrolysis and inactivation.Sulbactam resembles the penicillin derivatives in structure (Figure) and is able to preserve their activity by its ability to inhibit the action of beta-lactamases, particularly those of the Richmond classes II-V (gram-negative) and the group A beta-lactamases (gram-positive). Sulbactam is referred to as a “suicide inhibitor” because while forming an irreversible complex with the enzyme, it is destroyed in the process. By virtue of its ability to render the beta-lactamases inactive, sulbactam has been combined with ampicillin in an effort to restore its activity against a number of pathogens that have developed resistance by this mechanism.

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