Comparative Study of Phenotypic Detection Methods of Metallo-beta-lactamase (MβL) Among Carbapenem Resistant Gram-Negative Bacilli (GNB) Isolated from Intensive Care Units (IUC) Patients

2020 ◽  
Vol 29 (3) ◽  
pp. 75-80
Author(s):  
Raghda Hager ◽  
Bassant M. Sayed
Keyword(s):  
Intensive Care ◽  
Intensive Care Units ◽  
Carbapenem Resistance ◽  
Detection Methods ◽  
Diffusion Method ◽  
Beta Lactamase ◽  
Gram Negative ◽  
Carbapenem Resistant ◽  
High Prevalence ◽  
Synergy Test

Background: Metallo-beta-lactamase (MβL) mediated resistance is an emergency threat in health care settings, and its identification is essential for treatment and infection control. Objectives: this study aimed to detect the MβL prevalence in Gram negative bacilli (GNB) and to compare its phenotypic detection methods. Methodology: Ninety six (96) isolates of GNB were isolated from different clinical specimens collected from patients admitted to intensive care units (ICU) of Ain Shams Hospitals, from February 2018 to June 2018. Isolates were screened for carbapenem resistance with imipenem 10 µg and meropenem 10 µg discs. The resistant isolates were tested for antibiotic susceptibility by disc diffusion method, and Meropenem minimum inhibitory concentration (MIC) were determinated, then the production of MβL was detected by imipenem-ethylene diamine tetra-acetic acid (EDTA) combined disc test (IPM-EDTA CDT), ceftazidime -EDTA combined disc test (CAZ-EDTA CDT) and Imipenem - EDTA double disc synergy test (IPM-EDTA DDST).Results: Forty three (43) isolates (44.7%) were resistant to carbapenem. Klebsiella pneumoniae (K.pneumoniae) was the most common isolated species; 29 (67.4%) isolates. Forty (40) isolates (93%) were positive for MβL by IPM-EDTA CDT method, whereas 36 (83.7%) were positive by CAZ -EDTA CDT method and 19 isolates (44.2%) were positive for MβL by IPM-EDTA DDST. Conclusion: High prevalence of MβL was detected among our isolates and IMP-EDTA CDT can be used as a phenotypic test in detection of MβL production.

scholarly journals Molecular screening of carbapenemase-producing Gram-negative strains in Romanian intensive care units during a one year survey

2014 ◽  
Vol 63 (10) ◽  
pp. 1303-1310 ◽  
Author(s):  
Irina Gheorghe ◽  
Ilda Czobor ◽  
Mariana Carmen Chifiriuc ◽  
Elvira Borcan ◽  
Camelia Ghiţă ◽  
...  
Keyword(s):  
Intensive Care ◽  
Intensive Care Units ◽  
Rapid Evolution ◽  
Rapid Identification ◽  
Molecular Screening ◽  
Tissue Samples ◽  
Gram Negative ◽  
Carbapenem Resistant ◽  
High Prevalence ◽  
One Year

This is the first study, to our knowledge, performed on a significant number of strains (79 carbapenem-resistant Enterobacteriaceae and 84 carbapenem-resistant non-fermenting Gram-negative rods, GNRs) isolated from tissue samples taken from patients in the intensive care units of two large hospitals in Bucharest, Romania, between 2011 and 2012. The results revealed a high prevalence and great diversity of carbapenemase genes (CRG), in both fermenting and non-fermenting Gram-negative carbapenem-resistant strains. The molecular screening of carbapenem-resistant GNRs revealed the presence of worldwide-distributed CRGs (i.e. bla OXA-48 and bla NDM-1 in Enterobacteriaceae and bla OXA-23, bla VIM-4, bla OXA-10-like, bla OXA-60-like, bla SPM-like and bla GES-like in non-fermenting GNRs), reflecting the rapid evolution and spread of carbapenemase producers, particularly in hospitals. Rapid identification of the colonized or infected patients is required, as are epidemiological investigations to establish the local or imported origin of the respective strains.

PREVALENCE OF CARBAPENEM-RESISTANT ACINETOBACTER BAUMANNII (CRAB) IN MEDICAL AND SURGICAL INTENSIVE CARE UNITS (ICUS) OF JPMC, KARACHI

2019 ◽  
Author(s):  
Rabia Arshad
Keyword(s):  
Intensive Care ◽  
Antimicrobial Resistance ◽  
Acinetobacter Baumannii ◽  
Intensive Care Units ◽  
Carbapenem Resistance ◽  
Chronic Obstructive ◽  
Surgical Intensive Care ◽  
Carbapenem Resistant ◽  
Number Of Patients ◽  
Increased Risk

Background: Antimicrobial resistance is one of the research priorities of health organizations due to increased risk of morbidity and mortality. Outbreaks of nosocomial infections caused by carbapenem-resistant Acinetobacter Baumannii (CRAB) strains are at rise worldwide. Antimicrobial resistance to carbapenems reduces clinical therapeutic choices and frequently led to treatment failure. The aim of our study was to determine the prevalence of carbapenem resistance in A. baumannii isolated from patients in intensive care units (ICUs). Methods: This cross-sectional study was carried out in the Department of Microbiology, Basic Medical Sciences Institute (BMSI), Jinnah Postgraduate Medical Centre (JPMC), Karachi, from December 2016 to November 2017. Total 63 non-repetitive A. baumannii were collected from the patients’ specimens, admitted to medical and surgical ICUs and wards of JPMC, Karachi. The bacterial isolates were processed according to standard microbiological procedures to observe for carbapenem resistance. SPSS 21 was used for data analysis. Results: Out of the 63 patients, 40 (63.5%) were male. The age of the patient ranged from 15-85 year, with average of 43 year. 34.9% patients had been hospitalized for 3 days. Chronic obstructive pulmonary disease was present in highest number with average of 58.7% for morbidity. Number of patients on mechanical ventilation was highest (65.1%). All isolates were susceptible to colistin. The resistance to ampicillin-sulbactam, ceftazidime, ciprofloxacin, amikacin, piperacillin- tazobactam and meropenem was 82.5%, 81%, 100%, 87.3%, 82.5% and 82% respectively. Out of 82% CRAB, 77% were obtained from ICUs. Conclusion: This study has revealed the high rate of carbapenem resistance in A. baumannii isolates in ICUs thus leaving behind limited therapeutic options.

scholarly journals Prevalence of metallo-β-lactamase genes among Pseudomonas aeruginosa isolated from various clinical samples in China

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Wei Wang ◽  
Xiaoya Wang
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibiotic Susceptibility ◽  
Ethylenediaminetetraacetic Acid ◽  
Carbapenem Resistance ◽  
Opportunistic Pathogen ◽  
Detection Methods ◽  
Clinical Samples ◽  
Routine Practice ◽  
Carbapenem Resistant ◽  
High Prevalence

AbstractBackgroundPseudomonas aeruginosa is an opportunistic pathogen which is associated with nosocomial infections and causes various diseases including urinary tract infection, pneumonia, soft-tissue infection and sepsis. The emergence of P. aeruginosa-acquired metallo-β-lactamase (MBL) is most worrisome and poses a serious threat during treatment and infection control. The objective of this study was to identify antibiotic susceptibility, phenotypic detection of MBL production and to determine the prevalence of MBL genes in carbapenem-resistant P. aeruginosa isolated from different clinical samples.MethodsA total of 329 non-duplicate P. aeruginosa isolated from various clinical samples from two hospitals in China between September 2017 and March 2019 were included in this study. Phenotypic detection of MBL was performed by the combined detection method using imipenem and imipenem-ethylenediaminetetraacetic acid (EDTA) discs. MBL-encoding genes including blaVIM-1, blaVIM-2, blaIMP-1, blaIMP-2, blaSPM-1, blaSIM, blaNDM-1 and blaGIM were detected by polymerase chain reaction (PCR).ResultsOf the 329 P. aeruginosa, majority of the isolates were resistant to imipenem (77.5%) followed by meropenem (64.7%). Of the 270 P. aeruginosa isolates tested, 149 (55.2%) isolates were found to be positive for MBL detection. Of the different samples, 57.8% (n = 26) of P. aeruginosa isolated from blood were found to be positive for MBL production. Of the various MBL genes, blaIMP-1 (28.2%) was the most predominant gene detected followed by blaVIM-2 (18.8%), blaVIM-1 (16.1%), blaNDM-1 (9.4%), blaIMP-2 (6.7%), blaSIM (6.0%), blaSPM-1 (4.0%) and blaGIM (1.3%) genes.ConclusionsThe high resistance of P. aeruginosa toward imipenem and meropenem and the high prevalence of blaIMP-1 and blaVIM-2 set the alarm on the increasing, perhaps the increased, carbapenem resistance. In addition to routine antibiotic susceptibility testings, our results emphasize the importance of both the phenotypic and genotypic MBL detection methods in routine practice for early detection of carbapenem resistance and to prevent further dissemination of this resistant pathogen.

scholarly journals Genome-Wide Identification of Carbapenem-Resistant Gram- Negative Bacterial (CR-GNB) Isolates Retrieved From Hospitalized Patients in Bihar, India

2021 ◽  
Author(s):  
Namrata Kumari ◽  
Mukesh Kumar ◽  
Amit Katiyar ◽  
Abhay Kumar ◽  
Pallavi Priya ◽  
...  
Keyword(s):  
Resistance Genes ◽  
Carbapenem Resistance ◽  
Cross Sectional Study ◽  
Clinical Isolates ◽  
Cross Sectional ◽  
Gram Negative ◽  
Carbapenem Resistant ◽  
Genome Wide ◽  
Synergy Test ◽  
Global Threat

Abstract Carbapenemase-producing clinical isolates are becoming more common over the world, posing a severe public health danger, particularly in developing nations like India. Carbapenem-resistant Gram-negative bacterial (CR-GNB) infection has become a fast-expending global threat with limited antibiotic choice and significant mortality. The aim of this study was to highlight the carbapenem-resistance among clinical isolates of hospital admitted patients in Bihar, India. A cross-sectional study was conducted with 101 clinical isolates of E. coli, K. pneumoniae, A. baumannii, and P. aeruginosa. All GNB isolates were tested for their antimicrobial susceptibility using double disc synergy test / modified hodge test (DDST/MHT) and subsequently confirmed carbapenemase-producing isolates were evaluated for carbapenem-resistance genes using whole-genome sequencing (genotypically) method. The overall percentage of carbapenem-resistance among GNB was (17/101) 16.83%. The AMR analysis demonstrates a significantly high prevalence of blaCTX−M followed by blaSHV, blaTEM, blaOXA and blaNDM β-lactams carbapenem-resistance genes among clinical isolates of GNB. Co-occurrence of carbapenemase-encoding genes with blaNDM was found in 70.6% of carbapenemase-producing isolates. Our study highlights the mechanism of carbapenem-resistance to curb the overwhelming threat posed by emergence of drug-resistance in India.

scholarly journals Investigation of Metallo-Beta-Lactamases in Carbapenem Resistant Pseudomonas aeruginosa Strains by Phenotypic and Genotypic Methods

2020 ◽  
Vol 25 (3) ◽  
pp. 301-307
Author(s):  
M. Duygu Aksoy ◽  
H. Murat Tuğrul
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Ethylenediaminetetraacetic Acid ◽  
Carbapenem Resistance ◽  
Beta Lactamase ◽  
Bacterial Strains ◽  
Beta Lactamases ◽  
Carbapenem Resistant ◽  
Synergy Test ◽  
Phenotypic Tests ◽  
Positive Controls

Introduction: Carbapenem resistant Pseudomonas aeruginosa strains cause serious problems in treatment. A large number of identified metallo-beta-lactamase (MBL) enzymes produced by P. aeruginosa are one of the most important mechanisms in resistance to carbapenems. MBL genes are located on the chromosome or plasmid, and they can easily spread between different bacterial strains. The activities of these enzymes are zinc-dependent, and they are inhibited by ethylenediaminetetraacetic acid (EDTA). Therefore, this advantage is used in MBL identification tests. In this study, it was aimed to determine MBL among P. aeruginosa strains. Materials and Methods: MBL existence was investigated in 35 P. aeruginosa strains accepted to be mildly susceptible/resistant to any of the carbapenem group of antibiotics through phenotypic and genotypic methods. Phenotypic tests were performed as double disk synergy test (DDST), combined disk diffusion tests (CDDT) by using 0.1 M and 0.5 M EDTA, MBL E-test, and modified Hodge test (MHT). blaIMP, blaVIM, blaGIM, blaSIM, blaSPM genes and blaNDM gene were investigated by multiplex polimerase chain reaction (PCR) and PCR, respectively. Escherichia coli ATCC 25922 and P. aeruginosa ATCC 27853 standard bacteria were used in tests. VIM-1, VIM-2, IMP-13, SPM-1, NDM-1 type MBL-producing P. aeruginosa strains were used as positive controls. Results: Among the carbapenems resistant P. aeruginosa isolates, positivity of MBL was found as 54.2% by MBL E-test, 42.8% by DDST, 94.2% and 37.1% by CDDT method using 0.5 M and 0.1 M EDTA, respectively. Modified Hodge test and genotypic method did not detect MBL. Conclusion: In order to correctly evaluate the results of the phenotypic method, the investigation of resistance genes by molecular methods is also required. The most common metallo-beta-lactamase enzymes responsible for resistance to carbapenem in Pseudomonas were not observed. It was thought that different mechanisms might be responsible for the identified carbapenem resistance.

Predicting acquisition of carbapenem-resistant Gram-negative pathogens in intensive care units

2019 ◽  
Vol 103 (2) ◽  
pp. 121-127 ◽  
Author(s):  
L.F. Dantas ◽  
B. Dalmas ◽  
R.M. Andrade ◽  
S. Hamacher ◽  
F.A. Bozza
Keyword(s):  
Intensive Care ◽  
Intensive Care Units ◽  
Gram Negative ◽  
Carbapenem Resistant

Molecular Typing of Imipenem-ResistantAcinetobacter baumannii-calcoaceticusComplex in a Singapore Hospital Where Carbapenem Resistance Is Endemic

2007 ◽  
Vol 28 (8) ◽  
pp. 941-944 ◽  
Author(s):  
Thean Yen Tan ◽  
Karen Poh ◽  
Siew Yong Ng
Keyword(s):  
Tertiary Care ◽  
Carbapenem Resistance ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Care Hospital ◽  
Typing Method ◽  
Disk Diffusion Method ◽  
Carbapenem Resistant ◽  
High Prevalence ◽  
Resistant Strains

Objective.To investigate the molecular epidemiology of carbapenem-resistantAcinetobacter baumannii-calcoaceticuscomplex isolates in a tertiary care hospital where the prevalence of carbapenem resistance among these organisms is high.Design.The study was a prospective, observational study performed during an 8-month period (May 1 through December 31, 2004).A. baumanniiisolates recovered from all clinical samples during the study period were included in the study. Antibiotic susceptibility testing was performed using the disk diffusion method, and all carbapenem-resistant strains were typed by a polymerase chain reaction-based typing method.Setting.An 800-bed hospital in Singapore.Results.More than half of recovered isolates were clonally unrelated, with the remaining isolates grouped into 4 genotypes.Conclusions.The results of the study suggest that the high prevalence of carbapenem resistance amongAcinetobacterorganisms in this institution is not caused by the spread of a predominant clone and that other factors may need to be investigated.

scholarly journals Organ donor screening for carbapenem‐resistant gram‐negative bacteria in Italian intensive care units: the DRI n study

2019 ◽  
Vol 20 (1) ◽  
pp. 262-273
Author(s):  
Francesco Procaccio ◽  
Lucia Masiero ◽  
Francesca Vespasiano ◽  
Paolo A. Grossi ◽  
Carlo Gagliotti ◽  
...  
Keyword(s):  
Intensive Care ◽  
Intensive Care Units ◽  
Organ Donor ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Donor Screening ◽  
Carbapenem Resistant

Phenotypic Characterisation of Klebseilla pneumoniae Carbapenemase and Metallo Beta-lactamase in Carbapenem Resistant Gram-negative Bacteria

2021 ◽  
Author(s):  
Thresia . ◽  
Surya Sankar ◽  
Siju Joseph ◽  
V.R. Ambily ◽  
Anu Bosewell ◽  
...  
Keyword(s):  
Confirmatory Test ◽  
Diffusion Method ◽  
Gram Negative Bacteria ◽  
Beta Lactamase ◽  
Disc Diffusion ◽  
Beta Lactam ◽  
Disc Diffusion Method ◽  
Gram Negative ◽  
E Coli ◽  
Carbapenem Resistant

Background: Antibiotic resistance is an emerging concern in the therapy of clinical infections worldwide. Previous studies conducted in our laboratory have confirmed an increase in the prevalence of extended spectrum beta-lactamase (ESBL) among the Gram-negative bacterial pathogens associated with dogs, which could act as a potential source for the transfer of these resistant pathogens or their genetic determinants to human. Since carbapenems are the last resort drugs against these resistant pathogens, the study was aimed to isolate and characterise carbapenem resistance among Escherichia coli (E. coli), Klebsiella pneumoniae (K. pneumoniae) and Pseudomonas aeruginosa (P. aeruginosa) associated with common clinical infections in dogs.Methods: A total of 100 samples were collected from lesional skin, urine and anterior vagina of dogs presented to the Veterinary Hospitals of Kerala Veterinary and Animal Sciences University at Mannuthy and Thrissur. The samples were cultured onto Brain Heart Infusion Agar (BHIA), Eosin Methylene Blue (EMB) and Mac Conkey (MAC) for isolation of bacteria. Identification of the isolates was performed based on cultural, morphological and biochemical characteristics. The isolates were subjected to antimicrobial susceptibility test (ABST) against the 12 commonly used beta-lactam and non–beta-lactam group of antibiotics by disc diffusion method and further subjected to screening for ESBL double disc diffusion method. Carbapenem-resistant isolates were subjected to phenotypic confirmatory test for carbapenemase production employing Imipenem-EDTA and Ertapenem-boronic acid minimum inhibitory concentration (MIC) strip method.Result: Forty four Gram-negative bacterial isolates obtained were viz., E. coli (30), K. pneumonia (11) and P. aeruginosa (3) from the 100 samples. Apart from these, other isolates obtained were Staphylococcus spp. (53) and Bacillus spp. (2). All the Gram-negative isolates were subjected to ABST employing 12 common antibiotics belonging to beta-lactam and non-beta-lactam groups. Multidrug resistance (MDR) could be observed in 28 E. coli, 11 K. pneumoniae and three P. aeruginosa isolates. All the 42 MDR isolates showed positive results for ESBL production. A total of 14 isolates out of the 44 Gram-negative bacilli were found to be resistant to carbapenem either to imipenem, meropenem or ertapenem. Among the 14 Gram-negative isolates, nine turned out to be positive for metallo-beta-lactamase (MBL) and none for K. penumoniae carbapenemase (KPC) on phenotypic confirmatory test for detecting major carbapenemase enzymes. The present study documented that Gram- negative bacteria like E. coli, K. pneumoniae and P. aeruginosa isolated from dogs are showing an increase rate of resistance against carbapenems which are the last resort drugs against ESBL producers. Hence, there is an urgent need to curb the irrational and excessive use of antibiotics in veterinary sector.

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