scholarly journals New Delhi Metallo-β-lactamase (NDM)-mediated Carbapenem-Resistant Pseudomonas aeruginosa Clinical Isolates in Sudan

Author(s):  
Salma Elnour Rahma Mohamed ◽  
Alfadil Alobied ◽  
Mohamed Ibrahim Saeed ◽  
Wafa Mohamed Hussien

Carbapenem resistance mediated by NDM is particularly gruesome as this carbapenemase can hydrolyze a wide range of β-lactam antibiotics. Aim: This study aims to detect NDM mediated carbapenem resistance in clinical isolates of Pseudomonas aeruginosa. Materials and Methods: 50 multi-drug resistant clinical urinary isolates of Pseudomonas aeruginosa from three major hospitals in Khartoum state Sudan; Khartoum Teaching Hospital, Medical Army Hospital and Omdurman teaching hospital, in period from July 2016 to September 2017, were investigated for carbapenem resistance using standard disc diffusion method and underwent real-time PCR to detect carbapenem resistance gene blaNDM. Data were analyzed using IBM SPSS. Results: 60% were positive for the blaNDM, 82% were resistant to Imipenem and 75% of the samples were resistant to Meropenem. Conclusion: The emergence of carbapenem resistance is a global problem that requires earnest attention. To make the suitable preventive measures, the emergence of these genes must be monitored closely. Our findings revealed that carbapenem-resistant due to the gene blaNDM is accounted for 60% of the cases, and due to lack of proper data documentation about the emergence of this gene in Sudan, these cases to the best of our knowledge are the first to be reported in Sudan.

2021 ◽  
Vol 22 (4) ◽  
pp. 498-503
Author(s):  
A.O. Ettu ◽  
B.A. Oladapo ◽  
O.O. Oduyebo

Background: Pseudomonas aeruginosa has been highly associated with carbapenem resistance in which carbapenemases has been suggested to be a major contributory factor. Hence the objective of this study was to phenotypically detect KPC-type carbapenemase, metallo-β-lactamase and OXA-48 carbapenemase production in clinical isolates of P. aeruginosa in Lagos University Teaching Hospital (LUTH), NigeriaMethodology: One hundred and seventy-one P. aeruginosa isolates consecutively recovered from clinical specimens of patients with infections at the Medical Microbiology and Parasitology laboratory of the hospital were identified using MicrobactTM 24E kit. Preliminary screening for carbapenem resistance was determined by the disc diffusion method on Mueller-Hinton agar using single discs of meropenem and imipenem. Phenotypic detection of carbapenemase production among carbapenem-resistant isolates was performed by the combination disc test of meropenem-phenylboronic acid (MRPBO) and meropenem-dipicolinic acid (MRPDP) as recommended by EUCAST 2013 guideline. Results: Out of the 171 P. aeruginosa isolates, 35 (20.5%) were carbapenem non-susceptible (resistant) while carbapenemase production was detected in 27 (77.1%) of these carbapenem resistant isolates, and no enzyme was detected in 8 (22.9%). Of the 27 carbapenemase producing isolates, 22 (81.5%) produced MBL, 1 (3.7%) produced KPC, while 4 (14.8%) produced both KPC and MBL enzymes. Conclusion: This study revealed that carbapenem resistance among P. aeruginosa clinical isolates in our institution is gradually increasing. The mechanism for this rise is associated with carbapenemases, with MBL being the major carbapenemase involved. There is the need to ensure strict compliance with the LUTH infection control guidelines in order to check the rising incidence of infection caused by carbapenem resistant P. aeruginosa.   French title: Prévalence de la production de carbapénémases dans les isolats de Pseudomonas aeruginosa causant des infections cliniques à l'hôpital universitaire de Lagos, Nigéria   Contexte: Pseudomonas aeruginosa a été fortement associé à la résistance aux carbapénèmes dans laquelle les carbapénèmases ont été suggérées comme étant un facteur contributif majeur. Par conséquent, l'objectif de cette étude était de détecter phénotypiquement la production de carbapénémase de type KPC, de métallo-β-lactamase et de carbapénémase OXA-48 dans des isolats cliniques de P. aeruginosa au Lagos University Teaching Hospital (LUTH), Nigeria. Méthodologie: Cent soixante et onze isolats de P. aeruginosa récupérés consécutivement à partir d'échantillons cliniques de patients infectés au laboratoire de microbiologie médicale et de parasitologie de l'hôpital ont été identifiés à l'aide du kit MicrobactTM 24E. Le dépistage préliminaire de la résistance aux carbapénèmes a été déterminé par la méthode de diffusion sur disque sur gélose Mueller-Hinton en utilisant des disques uniques de méropénème et d'imipénème. La détection phénotypique de la production de carbapénèmes parmi les isolats résistants aux carbapénèmes a été réalisée par le test de disque combiné d'acide méropénème-phénylboronique (MRPBO) et d'acide méropénème-dipicolinique (MRPDP) tel que recommandé par la directive EUCAST 2013. Résultats: Sur les 171 isolats de P. aeruginosa, 35 (20,5%) étaient des carbapénèmes non sensibles (résistants) tandis que la production de carbapénèmes a été détectée dans 27 (77,1%) de ces isolats résistants aux carbapénèmes, et aucune enzyme n'a été détectée dans 8 (22,9%). Sur les 27 isolats producteurs de carbapénémases, 22 (81,5%) produisaient des MBL, 1 (3,7%) produisaient des KPC, tandis que 4 (14,8%) produisaient à la fois des enzymes KPC et MBL. Conclusion: Cette étude a révélé que la résistance aux carbapénèmes parmi les isolats cliniques de P. aeruginosa dans notre institution augmente progressivement. Le mécanisme de cette augmentation est associé aux carbapénémases, la MBL étant la principale carbapénémase impliquée. Il est nécessaire de garantir le strict respect des directives de contrôle des infections LUTH afin de contrôler l'incidence croissante des infections causées par P. aeruginosa résistant aux carbapénèmes.


2020 ◽  
Author(s):  
Doha Omer Ali ◽  
Mohamed M.A. Nagla

AbatractCarbapenem resistance in Pseudomonas.aeruginosa is particularly worrisome because this class of β-lactam represents the last therapeutic resource for control of bacterial infection.So this study aimed to detect the frequency of bla OXA-48 resistance gene among Pseudomonas aeruginosa clinical isolates during the period from November 2018 to November 2019.Hundred Pseudomonas aeruginosa clinical isolates, 81 carbapenems (imipenem meropenem) resistant and 19 carbapenems sensitive were collected from Omdurman Teaching Hospital, Fedail Hospital and Soba Teaching Hospital in Khartoum State-Sudan. All isolates were re-identified using conventional bacteriological techniques, their susceptibility to carbapenems were tested using Kirby-Bauer method for confirmation and investigated for the presence of the bla OXA-48 gene using conventional PCR technique.60 (60.0%) out of 100 Pseudomonas aeruginosa clinical isolates were positive for blaOXA-48 gene. Out of 81 carbapenem resistant isolates 54(66.7%) were positive for bla OXA-48 gene, while among the (19) carbapenem sensitive isolates 6 (31.6%) were positive for blaOXA-48 gene. There was statistically significant association between carbapenem resistant isolates and the presence of blaOXA-48 gene (P-value = 0.006).Wound swabs were the predominant clinical samples detected harboring bla OXA-48 gene both among the sensitive 5 (83.3%) and carbapenem resistant isolates 29(53.7) (P.value> 0.05).Our findings revealed high frequency of bla OXA-48 among carbapenem resistant isolates so identification of bla OXA-48 producing strains and taking efforts to reduce the rate of transferring these gene between the different strains is essential for optimization of therapy and improves of patients outcomes.


Author(s):  
Eucharia E. Nmema ◽  
Chioma S. Osuagwu ◽  
Eunice N. Anaele

Aims: The aims of the study were to evaluate the multidrug resistance profile and mechanisms of carbapenem resistance in Pseudomonas aeruginosa clinical isolates using phenotypic and genotypic methods. Study Design: A descriptive laboratory based study. Place and Duration of Study: Microbiology Laboratory, Ondo State University of Science and Technology, Okitipupa, and Biotechnology Laboratory, Ladoke Akintola University of Technology, Osogbo, Nigeria, between June 2017 and November 2018. Methodology: Ten P. aeruginosa isolates were recovered from patients at Lagos University Teaching Hospital, and susceptibilities to imipenem (10 µg), meropenem (10 µg) and a panel of antibiotics were performed by the disk diffusion method. Genotypic methods including Polymerase Chain Reactions (PCR) and agarose gel electrophoresis were carried out according to established protocols. oprD and blaIMP gene primers were used for the PCR amplification. Results: Fifty percent (50%) of the isolates showed multiple drug resistance. Four isolates (40%) were carbapenem resistant (CR). oprD gene was detectedin 90% (9/10) of the isolates. 75% (3/4) of CR strains were among the strains showing oprD gene. 25% (1/4) CR strain (PA1421) was oprD negative. Loss or mutation of oprD gene seems to be the mechanism of carbapenem resistance in strain PA1421. Conclusion: Loss or mutation of oprD gene was identified in this study as a mechanism of carbapenem resistance. oprD gene encodes the outer membrane protein (OprD) porin in P. aeruginosa whose deficiency confers resistance to carbapenems, especially imipenem. Surveillance of the antimicrobial susceptibility patterns of P. aeruginosa is of critical importance in understanding new and emerging resistance trends, reviewing antibiotic policies and informing therapeutic options.


2018 ◽  
Vol 11 (12) ◽  
pp. 935-943 ◽  
Author(s):  
Mona Shaaban ◽  
Ahmed Al-Qahtani ◽  
Mohammed Al-Ahdal ◽  
Rasha Barwa

Introduction: Emergence of carbapenem resistance in Pseudomonas aeruginosa increases the therapeutic dilemma. In this study, we investigated various mechanisms involved in the resistance of P. aeruginosa clinical isolates to carbapenems. Methodology: P. aeruginosa isolates were isolated from different clinical samples. The antimicrobial susceptibility was evaluated by disc diffusion method. Carbapenemases were detected among carbapenem resistant isolates. Expression level of mexB and oprD was determined by real-time PCR. Molecular relatedness among isolates was detected based on pulse-field gel electrophoresis (PFGE). Results: Ninety P. aeruginosa isolates were purified from clinical specimens. High levels of resistance to imipenem and meropenem were detected in 16 isolates. PCR analysis of carbapenemases indicated the prevalence of Verona integron-encoded metallo-beta-lactamase (VIM); four isolates produced only VIM enzymes (VIM-1 or VIM-2), while the remaining twelve co-produced both VIM-1 or VIM-2 and NDM enzymes. Additionally, real-time PCR analysis elucidated high expression levels of mexB in seven of the carbapenem resistant isolates and low expression of oprD in seven isolates. The identified carbapenem-resistant isolates were clustered into eleven PFGE profiles where clusters E1 and E2 involved isolates exhibiting multiple carbapenemase genes (blaNDM-1, blaVIM-1 and blaVIM-2). Conclusion: Various mechanisms underlying carbapenem resistance have been detected in our P. aeruginosa cohort of isolates. Emergence of P. aeruginosa as a reservoir of multiple carbapenemases is increasing over time limiting the treatment options to this serious infection. This increases the urgency for infection control practices to reduce the incidence of this infection.


2001 ◽  
Vol 45 (2) ◽  
pp. 480-484 ◽  
Author(s):  
Hyunjoo Pai ◽  
Jong-Won Kim ◽  
Jungmin Kim ◽  
Ji Hyang Lee ◽  
Kang Won Choe ◽  
...  

ABSTRACT In order to define the contributions of the mechanisms for carbapenem resistance in clinical strains of Pseudomonas aeruginosa, we investigated the presence of OprD, the expressions of the MexAB-OprM and MexEF-OprN systems, and the production of the β-lactamases for 44 clinical strains. All of the carbapenem-resistant isolates showed the loss of or decreased levels of OprD. Three strains overexpressed the MexAB-OprM efflux system by carrying mutations inmexR. These three strains had the amino acid substitution in MexR protein, Arg (CGG) → Gln (CAG), at the position of amino acid 70. None of the isolates, however, expressed the MexEF-OprN efflux system. For the characterization of β-lactamases, at least 13 isolates were the depressed mutants, and 12 strains produced secondary β-lactamases. Based on the above resistance mechanisms, the MICs of carbapenem for the isolates were analyzed. The MICs of carbapenem were mostly determined by the expression of OprD. The MICs of meropenem were two- to four-fold increased for the isolates which overexpressed MexAB-OprM in the background of OprD loss. However, the elevated MICs of meropenem for some individual isolates could not be explained. These findings suggested that other resistance mechanisms would play a role in meropenem resistance in clinical isolates of P. aeruginosa.


2020 ◽  
Vol 41 (S1) ◽  
pp. s398-s398
Author(s):  
Tuan Huynh ◽  
Vasquez Amber ◽  
Lan Pham ◽  
Loan Luong ◽  
Tuan Le ◽  
...  

Background: Carbapenem-resistant gram-negative bacteria are an urgent threat to healthcare safety around the world. In Vietnam, Although surveillance and control of multidrug-resistant organisms is a national priority, information on the burden of these resistant pathogens is still scarce. At University Medical Center Ho Chi Minh City, Vietnam, we aimed to better understand carbapenem-resistance through 2 phases: (1) assess proportion of carbapenem-resistant gram-negative organisms that are carbapanemase-producing (CP-CRO) and (2) assess transmission burden of carbapenemase-producing carbapenem-resistant Enterobacterieacea (CP-CRE) in the general intensive care unit (ICU). Methods: In the first phase, all gram-negative clinical isolates collected between November 2018 and April 2019 were tested for carbapenem-resistance using the disc-diffusion method and were defined as meropenem resistant using the Clinical and Laboratory Standards Institute 2018 break point (M100-Performance Standards for Antimicrobial Susceptibility Testing, 28th Edition). Carbapenem-resistant bacteria were tested for phenotypic carbapenemase-production using the Becton Dickinson Phoenix CPO Detect assay. In the second phase, we instituted CP-CRE rectal screening using CHROMagar mSuperCARBA media for all ICU patients from July through September 2019. Patients were screened on admission, and negative patients were rescreened every 2 days until discharge, death, or CRE-positive screening or culture. Admission prevalence and incidence of CP-CRE transmission was calculated among CP-CRE infected or colonized patients. Results: From November 2018 through April 2019, 599 gram-negative clinical isolates from 543 patient samples were identified. Of these, 108 were carbapenem-resistant; 107 (99%) of carbapenem-resistant isolates were carbapenemase-producing by phenotypic method. Most CP-CRO were Acinetobacter baumannii (45 of 107, 42%) or Klebsiella pneumoniae (39 of 107, 36%). During ICU CP-CRE colonization screening, the July positivity rate on admission was 40% (32 of 81), the August positivity rate on admission was 30% (21 of 71), and the September positivity rate on admission was 40% (30 of 75). Of those with negative admission screen, the proportion of new CP-CRE colonization in July was 45% (22 of 49), the proportion of new CP-CRE colonization in August was 64% (32 of 50), and the proportion of new CP-CRE colonization in September was 44% (20 of 45). Across all 3 months of screening, the proportions of CP-CRE that were Klebsiella, Citrobacter, or Enterobacter were 68% (118 of 174) and the proportion of CP-CRE that were Eschericia coli was 37% (56 of 174). The average number of days to turn from negative to positive screening result was 4.1. Conclusions: Our analysis demonstrates that nearly all carbapenem-resistant organisms at our hospital are carbapenemase producing. In the ICU, we identified a high burden of CP-CRE, attributable to high presence on admission and new acquisition in the ICU. An intervention package based on CDC-recommended enhanced infection control measures is being implemented to decrease CP-CRE transmission in the ICU.Funding: NoneDisclosures: None


2006 ◽  
Vol 50 (5) ◽  
pp. 1633-1641 ◽  
Author(s):  
John Quale ◽  
Simona Bratu ◽  
Jyoti Gupta ◽  
David Landman

ABSTRACT Carbapenems are important agents for the therapy of infections due to multidrug-resistant Pseudomonas aeruginosa; the development of carbapenem resistance hampers effective therapeutic options. To assess the mechanisms leading to resistance, 33 clinical isolates with differing degrees of carbapenem susceptibility were analyzed for the expression of the chromosomal β-lactamase (ampC), the porin that is important for the entry of carbapenems (oprD), and the proteins involved in four efflux systems (mexA, mexC, mexE, and mexX). Real-time reverse transcriptase PCR was performed using primers and fluorescent probes for each of the target genes. The sequencing of regulatory genes (ampR, mexR, nalC, nalD, mexT, and mexZ) was also performed. Diminished expression of oprD was present in all imipenem- and meropenem-resistant isolates but was not required for ertapenem resistance. Increased expression of ampC was not observed in several isolates that were overtly resistant to carbapenems. Increased expression of several efflux systems was observed in many of the carbapenem-resistant isolates. Increased efflux activity correlated with high-level ertapenem resistance and reduced susceptibility to meropenem and aztreonam. Most isolates with increased expression of mexA had mutations affecting nalC and/or nalD. Two isolates with mutations leading to a premature stop codon in mexZ had markedly elevated mexX expressions, although mutations in mexZ were not a prerequisite for overexpression. β-Lactam resistance in clinical isolates of P. aeruginosa is a result of the interplay between diminished production of oprD, increased activity of ampC, and several efflux systems.


2021 ◽  
Vol 18 (4) ◽  
pp. 429-436
Author(s):  
Santhiya K. ◽  
Jayanthi S. ◽  
Ananthasubramanian M. ◽  
Appalaraju B.

Background: Carbapenem-resistant Enterobacteriaceae (CRE) has emerged as a global threat with mortality risk ranging from 48%-71% worldwide. The emergence of MBL resistance is threatening as carbapenem is one of the last line antibiotics. A total 24 variants of NDM resistance raises a concern to the clinicians and epidemiologists worldwide. Objective: The study aims at identifying MBL resistance (NDM, IMP, VIM, GIM, SPM, and SIM) and its coexistence in clinical isolates in a single tertiary care center. Methodology: Forty five clinical isolates characterized phenotypically for Carbapenem resistance obtained from PSG Institute of Medical Science and Research (PSG IMSR), Coimbatore, between February to March 2018 were taken for analysis. Result: Out of the 45 Clinical isolates, 38 isolates (84%) were detected as MBL carriers. VIM, NDM, GIM, and SPM were the predominant resistance genes, with detection rates of 48.8%, 28.8%, 24.4%, and 22.2% respectively. Fifteen isolates were observed to harbor more than one MBL gene in coexistence. Two isolates - U42 and R714 (K. pneumoniae) were found to harbor all 5 MBL variants in combination. Conclusion: 33% of clinical isolates harboring multiple MBL variants is a concern in clinical settings. The presence of SPM and GIM gene amongst isolates in this geographical location within India is an indicator demanding continuous monitoring of these resistance determinants.


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