Detection and characterization of carbapenem resistant Gram‐negative bacilli isolates recovered from hospitalized patients at Soba University Hospital, Sudan (original) (raw)
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Research Square (Research Square), 2020
Background: Antimicrobial resistance (AMR) poses a threat to global health security. Whilst over the past decade, there has been an increase in reports of nosocomial infections globally caused by carbapenem resistant Gram-negative bacilli (GNB). This study aimed to detect and characterize carbapenem resistance Gram negative bacteria isolated from hospitalized patients in Soba University Hospital (SUH) in Khartoum State, Sudan Results: A total of 206 GNB isolates from different clinical specimens were obtained from hospitalised patients between October 2016 to February 2017. Of 206 isolates, 171 (83%) were con rmed resistant phenotypically and 121 (58.7%) isolates were positive for the presence of one or more carbapenemase genes. New Delhi metallo-β-lactamase (NDM) types were the most predominant genes, blaNDM 107(52%). Others included blaIMP 7 (3.4%), blaOXA-48 5(2.4%), blaVIM 2 (0.9%) and blaKPC 0 (0%). Coresistance genes with NDM producing GNB were detected in 87 (81.3%) of all blaNDM positive isolates. A signi cant association between phenotypic and genotypic resistance was observed (P < 0.001). NDM1 was the most frequent subtype observed in 75 (70 %) isolates which clusters to the Indian lineage. Conclusions: The frequency of carbapenemase producing bacilli was found to be improperly high in SUH. NDM was found to be the most prevalent carbapenemase gene among clinical isolates. Close surveillance across all hospitals in Sudan is required. The relative distribution of carbapenemase genes among GNB in nosocomial infections in Africa needs to be de ned.
2019
Background Carbapenems are broad-spectrum β-lactam antibiotics widely prescribed for the treatment of multidrugresistant gram negative bacilli in systemic infections. In the past ten years the Carbapenem resistance among gram-negative bacilli is rapidly expanding across nosocomial infection isolates. This study was conducted to determine the prevalence and to characterize Carbapenem resistance genes among Gram negative bacilli (GNB) isolated from patients treated in hospitals in Khartoum state, Sudan. Methods A cross-sectional laboratory based study was conducted over six months at the microbiology department
International journal of science and research, 2024
Background: Multi-drug resistance among gram negative bacteria with special interest to carbapenem resistance has been increasingly noticed worldwide leaving very few treatment options and associated with high morbidity and mortality. The rapid and accurate detection of several carbapenemase genes in these bacteria is important for both clinicians and infection control practitioners. In this work, we used a multiplex PCR assay for simultaneous detection of 5 carbapenemase genes among GNBs in a single run within a short time. Objectives: Use of Multiplex PCR for early detection of plasmid-borne carbapenemase genes among carbapenem resistant gram-negative bacteria isolated from various clinical samples of patients in community and hospital. Methods: This observational study was conducted at microbiology and molecular laboratory of a Tertiary care hospital in Dhaka, Bangladesh from June to July, 2023. Fresh culture colonies of 30 Carbapenem resistant and 04 Carbapenem sensitive gram-negative bacteria from different clinical samples were tested for carbapenem resistance by both Kirby-bauer disc diffusion and automated MIC detection as per CLSI guidelines. A multiplex PCR assay was done with Unimedica Multiplex Real time PCR Kit for identification of KPC, NDM, VIM, IMP and OXA-48 Carbapenem Resistance Genes and results were analyzed by software. Results: Among total tested 34 clinical isolates, 16 were Klebsiella pneumoniae, 04 E. coli, 08 Pseudomonas aeruginosa and 06 Acinetobacter baumannii. Of them, 24(71%) MDR-GNBs showed the presence of NDM and OXA-48 gene on Multiplex PCR. Both NDM and OXA-48 were co expressed predominantly in 50% isolates, while 33.3% NDM and 16.7% OXA-48 were detected solitarily. No KPC, VIM, IMP were determined. Minocycline (50%), tigecycline, fosfomycin and gentamicin (30%) & cotrimoxazole (25%) were sensitive for NDM encoded carbapenem resistant GNBs, however no sensitivity found to ceftazidimeavibactam. OXA-48 harbouring CR-GNBs showed 25% Fosfomycin & Ceftazidime-avibactam sensitivity and 100% resistance to all other tested antibiotics. Combined NDM & OXA 48 genes were positive in 60% K. pneumoniae, 50% E coli and 28.60% Pseudomonas aeruginosa. They were 33% sensitive to tigecycline, minocycline & fosfomycin, 17% to gentamicin and 8% to ceftazidime-avibactam & cotrimoxazole. All isolates were 100% sensitive to colistin and polymyxin B. Though having high MIC, no resistance genes were present in 6 carbapenem resistant Acinatobacter baumannii. Conclusion: Multiplex PCR overcome the limitations of the phenotypic methods and automated systems in identification of carbapenemase genes that enable physicians to select the most appropriate antibiotics. Our study has shown the coexistence of multiple genes in a single bacteria pointing out that different carbapenmases enzymes are utilized by the bacteria to inactive the carbapenem drugs. We recommend routine testing for carbapenem resistance genes among the MDR-GNB infections which will contribute in preserving carbapenems, the last resort antibiotics.
American Journal of Infectious Diseases and Microbiology, 2021
Carbapenem-resistant Enterobacteriaceae strains have been responsible for an increasing number of hospital-acquired infections globally. This study aimed to determine the prevalence of carbapenemase-producing Enterobacteriaceae and their frequency of antimicrobial-resistant patterns among hospitalized patients across three Khartoum State Teaching Hospitals, Sudan. Materials and Methods. A cross-sectional study was conducted at Khartoum State Teaching Hospitals from April 2018 to October 2019. A total of 384 non-duplicative Enterobacteriaceae strains were isolated from1062 clinical samples obtained from hospitalized patients receiving treatment across three main teaching hospitals. The samples were cultured into a MacConkey agar plate. The Enterobacteriaceae strains were differentiated by specific colony color and again by biochemical test. Antimicrobial susceptibility testing was performed by the disc diffusion method. The minimum inhibitory concentration (MIC) of imipenem and meropenem was performed by the agar dilution method. Multiplex polymerase chain reaction (PCR) was performed to investigate the presence of carbapenemase-encoding genes. Data analysis was carried out using SPSS version 21. Results. Of the 36.2% (384/10.62) nonduplicate of Enterobacteriaceae strains isolated from clinical samples, 122 (31.8%) were carbapenemase-producing Enterobacteriaceae (CPE). Of these isolates, 37 (30.3%) harbored the blaIMP followed by; 29 (23.8%) blaNDM, 21 (17.2%) blaOXA-48, 6 (4.9%) blaGES, 5 (4.1%) were blaKPC, 3(2.5%) blaGIM-1, 2(1.6%) blaVIM and 1 (0.8%) blaSIM-1, while the remaining 19(15.6%) isolates carried combinations carbapenemase blagenes. The most predominant CPE strains were Escherichia coli 40 (32.8%) followed by Klebsiella pneumoniae 24 (19.7%) and Enterobacter aerogenes 14(11.5%). Most of the CPE isolates were isolated from wound swab 40(32.8), sputum 33(27.0), and urine 22(18.0) samples. Furthermost strains showed high resistance rates (>70%) to the antibiotics tested. Resistance to amikacin, tetracycline, co-trimoxazole, and nalidixic acid was 36.9%, 43.4%, 62.3%, and 63.9%, respectively and 82.8 % of CPE strains were susceptible to colistin. The detection of blagenes carbapenemases in CPE strains had a significant effect on both imipenem and meropenem MICs. Conclusion. The most prevalent carbapenemase-producing blagenes among clinical Enterobacteriaceae clinical isolates from the three Khartoum state regions were blaIMP, blaNDM, and blaOXA-48. In contrast, the propensity of the multidrug-resistant profile that has been associated with producing carbapenemase blagenes is alarming. Therefore, it is very important to establish a routine screening of carbapenemase-producing blagenes in clinical isolates to prevent the dissemination of resistant strains among both inpatients and outpatients in hospital settings.
PLOS ONE, 2021
Background Emerging worldwide in the past decade, there has been a significant increase in multidrug-resistant bacteria from serious nosocomial infections, especially carbapenemase-producing Gram-negative bacilli that have emerged worldwide. The objective of this study is to investigate carbapenem resistance in Gram-negative bacilli bacteria using phenotypic detection, antimicrobial resistance profiles and genotypic characterisation methods. Methods 200 Gram-negative bacilli isolates were collected from different clinical specimens. All clinical samples were exposed to isolation and identification of significant pathogens applying bacteriological examination and an automated Vitek-2 system. The isolates were subjected to susceptibility tests by the Vitek-2 automated system and those isolates that were resistant to beta-lactam drugs, including carbapenems, third-generation cephalosporines or cefoxitin, were selected for phenotyping using Carba plus disc system assay for detection of ...
Antibiotics
Carbapenem resistance (CR) is an emerging health issue. Epidemiological surveys on carbapenem-resistant Gram-negative bacilli (CR-GNB) in Lebanon remain scarce. In this study, we determined the prevalence of CR-GNB isolated between 2015 to 2019 in three hospitals in northern Lebanon: 311 CR-Enterobacterales (out of 11210; 2.8%), 155 CR-Pseudomonas (out of 1034; 15%) and 106 CR- Acinetobacter (out of 184; 57.6%) were identified. CR mechanisms were determined for 146 randomly chosen isolates: the Carba NP test revealed an enzymatic resistance to carbapenems in 109 isolates (out of 146, 74.7%). Produced carbapenemases were evaluated by the NG-Test Carba5, NG-Test OXA-23 immunochromatographic assays and PCR. Carbapenemase-producing (CP) Enterobacterales expressed blaOXA-48-like, blaNDM-like and blaVIM-like genes and CP-Pseudomonas expressed blaIMP-like and blaVIM-like genes, whereas CP-Acinetobacter expressed blaOXA-23-like genes. The NG-Test Carba5 results were confirmed by PCR sequenc...
F1000Research
Background: Carbapenems are used as antibiotics of last resort for treating infections due to multidrug-resistant Gram-negative bacilli, but emergence of Carbapenem resistant Gram-negative bacilli have been reported due to the production of Carbapenemase enzymes that significantly limits treatment options for life-threatening infections. Objective: This study aimed to detect Carbapenem resistant Gram-negative bacilli from patients attended to different hospitals in Khartoum state and to detect Carbapenemase enzymes production by phenotypic and genotypic methods. Methods: A hospital based cross sectional study was conducted in Khartoum state in the period from February to August 2016. Hundred and forty nine Gram-negative bacilli bacteria were isolated from different clinical specimens. Blood agar, Chromogenic agar media, MacConkey agar, XLD mediaandstandard biochemical tests were used for isolation and identification of Gram-negative bacilli from different samples. Standard antimicro...
BioMed Research International, 2014
The burden of antimicrobial resistance (AMR) is rapidly growing across antibiotic classes, with increased detection of isolates resistant to carbapenems. Data on the prevalence of carbapenem resistance in developing countries is limited; therefore, in this study, we determined the prevalence of carbapenemase genes among multidrug resistant gram negative bacteria (MDR-GNB) isolated from clinical specimens in a tertiary hospital in Mwanza, Tanzania. A total of 227 MDR-GNB isolates were analyzed for carbapenem resistance genes. For each isolate, five different PCR assays were performed, allowing for the detection of the major carbapenemase genes, including those encoding the VIM-, IMP-, and NDM-type metallo-beta-lactamases, the class A KPC-type carbapenemases, and the class D OXA-48 enzyme. Of 227 isolates, 80 (35%) were positive for one or more carbapenemase gene. IMP-types were the most predominant gene followed by VIM, in 49 (21.59%) and 28 (12%) isolates, respectively. Carbapenemase genes were most detected in K. pneumoniae 24 (11%), followed by P. aeruginosa 23 (10%), and E. coli with 19 isolates (8%). We have demonstrated for the first time a high prevalence of MDR-GNB clinical isolates having carbapenem resistance genes in Tanzania. We recommend routine testing for carbapenem resistance among the MDR-GNB particularly in systemic infections. VIM , IMP , KPC , and NDM , respectively [6]. Recently, increasing resistance to carbapenems in health care associated infections has been reported worldwide . Studies in New York City found 39% of patients with fecal colonization of KPC producing K. pneumoniae . In Africa, data on the prevalence and distribution of carbapenem
Egyptian Academic Journal of Biological Sciences, G. Microbiology
Background: Antimicrobial resistance is a great scourge on human health, exacerbated by the acquisition of resistance to carbapenems, the last resort treatment for infections caused by extended-spectrum beta-lactamase (ESBL) producing bacteria. Objectives: This cross-sectional study evaluated the prevalence of genes encoding ESBL and carbapenemase production in Gramnegative bacteria from clinical samples in Lagos state, Nigeria.Method: A total of 107 bacteria cultures were obtained from hospitals and clinical diagnostic laboratories. Isolate identification, antibiotics susceptibility testing, and phenotypic detection of ESBL production were done using standardized procedures. Multiplex polymerase chain reaction (PCR) was performed on ESBL-producing isolates to detect blaTEM, blaSHV, blaCTX-M, blaKPC, blaVIM, blaIMP and blaOXA.Result: Among the 107 cultures, 83 isolates were obtained with 55 being Gram-negative. Escherichia coli (22; 40%) was the most prevalent species followed by Klebsiella pneumoniae (7; 13%). Multidrug resistance (MDR) was observed in 34 (62%) of the isolated bacteria with 14 (26%) not susceptible to meropenem. ESBL production was detected in 42 (76%) of the isolates of which 23 (55%) strains harboured one or more of the genes blaTEM, blaSHV, and blaCTX-M. The carbapenemase genes blaKPC and/or blaVIM were observed in 11 (26%) isolates. No isolated bacteria were found to harbour blaIMP and/or blaOXA.Conclusion: Genes encoding ESBL and carbapenemase production were detected in samples of human origin in Lagos state. Novel antibiotics and/or alternative therapy are necessary for infection therapy in the near future.
Journal of Global Antimicrobial Resistance, 2017
Objectives: The aim of this study was to investigate the molecular epidemiology and the genetic support of carbapenem-resistant Enterobacteriaceae and Acinetobacter spp. isolates collected in the University Hospital of Ouargla, southern Algeria. Methods: A total of 99 Gram-negative bacteria (GNB) were collected from stool samples of colonised patients and from inanimate surfaces in the hospital environment between December 2014 and August 2015. Selected Enterobacteriaceae and Acinetobacter spp. isolates with reduced susceptibility to carbapenems were subjected to phenotypic study, including antibiotic susceptibility testing according to CA-SFM-EUCAST 2015 guidelines and modified Carba NP test. Genes encoding carbapenemases, extendedspectrum -lactamases (ESBLs) and AmpC -lactamases were screened by PCR and sequencing. Clonal relatedness was determined by multilocus sequencing typing (MLST). Results: Of the 99 GNB isolates, 10 (10.1%) showed reduced susceptibility to carbapenems were studied further, including 7 Acinetobacter baumannii, 1 Acinetobacter nosocomialis, 1 Escherichia coli and 1 Klebsiella pneumoniae. PCR and sequencing showed that four A. baumannii isolates and the single A. nosocomialis isolate harboured bla NDM-1. In addition, bla OXA-23 was observed in three A. baumannii isolates, and bla OXA-48 was detected in the two Enterobacteriaceae isolates. MLST assigned the K. pneumoniae to ST999 and the E. coli to ST38. The seven A. baumannii isolates belonged to ST85 and ST2. Conclusions: This study describes the epidemiology of carbapenemases produced by Enterobacteriaceae and Acinetobacter spp. in southern Algeria and reports the first description of metallo--lactamase NDM-1-producing A. nosocomialis in Algeria.