scholarly journals Prevalence of multidrug-resistance and bla and blaIMP genes among gram-negative clinical isolates in tertiary care hospital, Kathmandu, Nepal

Author(s):  
Mehraj Ansari ◽  
Subhas Chandra Aryal ◽  
Ganesh Rai ◽  
Kul Raj Rai ◽  
Susil Pyakurel ◽  
...  

Background and Objectives: Carbapenems have been the choice of antibiotics for the treatment of infections caused by multidrug-resistant bacteria. The main objective of this study was to determine the prevalence of carbapenemase (blaVIM and bla ) producing isolates among Enterobacteriaceae, Pseudomonas aeruginosa, and Acinetobacter baumannii. Materials and Methods: A total of 1,151 clinical samples were collected from the patients visiting Annapurna Neurological Institute and Allied Science and Annapurna Research Centre, Kathmandu, between June 2017 and January 2018. Antibiotic susceptibility testing (AST) was performed on the Enterobacteriaceae, P. aeruginosa and A. baumannii isolates using the Kirby-Bauer disk diffusion method. The modified Hodge test (MHT) was performed on the carbapenem-resistant isolates to confirm carbapenemase production. DNA was extracted and then screened for blaVIM and blaIMP genes by multiplex PCR. Results: Of the total 1,151 clinical samples, 253 (22.0%) showed positive growth. Of them, 226 (89.3%) were identified as Enterobacteriaceae, P. aeruginosa, and A. baumannii. Among the 226 isolates, 106 (46.9%) were multidrug-resistant. Out of the 106, 97 (91.5%) isolates showed resistance to at least one of the carbapenem used. Among the 97 carbapenem-resistant isolates, 67 (69.1%) showed the modified Hodge test (MHT) positive results. bla isolates respectively using multiplex PCR assay. Conclusion: This study determined a high prevalence of MDR and carbapenem resistance among Enterobacteriaceae, P. aeruginosa, and A. baumannii as detected by the presence of blaVIM and blaIMP genes. This study recommends the use of rapid and advanced diagnostic tools along with conventional phenotypic detection methods in the clinical settings for early detection and management of drug-resistant pathogens to improve treatment strategies.

2021 ◽  
Vol 23 (4) ◽  
pp. 290-296
Author(s):  
Rojina Darnal ◽  
Mehraj Ansari ◽  
Ganesh Rai ◽  
Kul Raj Rai ◽  
Shiba Kumar Rai

Carbapenemases are the enzymes that catalyze β–lactam groups of antibiotics. The carbapenemase producers are resistant to β–lactam antibiotics and are usually multidrug-resistant bacteria challenging widely used therapeutics and treatment options. Therefore, the detection of carbapenemase activity among clinical isolates is of great therapeutic importance. We aimed to study the MDR and carbapenemase-producing Klebsiella pneumoniae and Pseudomonas aeruginosa isolated from various clinical samples at a tertiary care hospital in Nepal. A total of 3,579 clinical samples were collected from the patients visiting the Department of Microbiology, B&B Hospital, Gwarko, Lalitpur. The samples were processed to isolate K. pneumoniae and P. aeruginosa and then subjected to antibiotic susceptibility testing (AST) by the Kirby-Bauer disk diffusion method. Phenotypic detection of carbapenemase activity was performed in the imipenem-resistant isolates by the modified Hodge test (MHT). Of the total samples, 1,067 (29.8%) samples showed significant growth positivity, out of which 190 (17.3%) isolates were K. pneumoniae and 121 (11.3%) were P. aeruginosa. Multidrug resistance was seen in 70.5% of the K. pneumoniae isolates and 65.3% of the P. aeruginosa isolates. Carbapenemase production was confirmed in 11.9%, and 12.2% of the imipenem-resistant K. pneumoniae and P. aeruginosa isolates, respectively, by the MHT. This study determined the higher prevalence of MDR among K. pneumoniae and P. aeruginosa; however, carbapenemase production was relatively low.


2019 ◽  
Vol 11 (03) ◽  
pp. 206-211
Author(s):  
Jaison Jayakaran ◽  
Nirupa Soundararajan ◽  
Priyadarshini Shanmugam

Abstract INTRODUCTION: Urinary tract infections (UTIs) remain as the most common infection. Catheter-associated (CA) UTI can lead to bacteremia and thereby is the leading cause of morbidity and mortality in hospitalized patients in our country. AIMS AND OBJECTIVES: This study aims to check the prevalence of CAUTI and study the phenotypic and genotypic characters of the multidrug-resistant organisms in a tertiary care hospital, with special reference to NDM-1 and OXA-23. MATERIALS AND METHODS: A total of 231 urine samples from patients with CA-UTI in different wards in a tertiary care hospital over a period of 3 months between June and August 2018 were collected and processed following the standard protocol. Antibiotic susceptibility tests were performed by disk-diffusion method. Modified Hodge test (MHT) was done to isolate carbapenem-resistant isolates, and polymerase chain reaction was done to detect NDM-1 and OXA-23. RESULTS: Out of 231 samples, 101 samples yielded significant growth. These 38 samples were Gram-negative bacilli which were resistant to carbapenems. Out of the 38 which showed carbapenem resistance, 23 were MHT positive. Out of the 23 MHT-positive isolates, 8 (21.05%) were positive for NDM-1 gene and only 1 (2.6%) was positive for the OXA-23 gene. CONCLUSION: This study has shown that carbapenem-resistant isolates from all the CA urinary tract-infected patients were 52.77% and most of them were Klebsiella. About 21% of them harbored the NDM-1 gene whereas only 2% had the OXA-23 gene. There has been an alarming increase in the spread of carbapenem resistance.


2021 ◽  
Vol 49 (1) ◽  
Author(s):  
Susil Pyakurel ◽  
Mehraj Ansari ◽  
Smriti Kattel ◽  
Ganesh Rai ◽  
Prasha Shrestha ◽  
...  

Abstract Aim Although carbapenem is the last-resort drug for treating drug-resistant Gram-negative bacterial infections, prevalence of carbapenem-resistant bacteria has substantially increased worldwide owing to irrational use of antibiotics particularly in developing countries like Nepal.  Therefore, this study was aimed to determine the prevalence of carbapenemase-producing K. pneumoniae and to detect the carbapenemase genes (blaNDM-2 and blaOXA-48) in at a tertiary care hospital in Nepal. Materials and methods A hospital-based cross-sectional study was carried out from June 2018 to January 2019 at the Microbiology Laboratory of Annapurna Neurological Institute and Allied Sciences, Kathmandu, Nepal. Different clinical samples were collected and cultured in appropriate growth media. Biochemical tests were performed for the identification of K. pneumoniae. Antibiotic susceptibility testing (AST) was performed by the Kirby–Bauer disc diffusion method. The modified Hodge test (MHT) was performed to detect carbapenemase producers. The plasmid was extracted by the modified alkaline hydrolysis method. Carbapenemase-producing K. pneumoniae were further confirmed by detecting blaNDM-2 and blaOXA-48 genes by PCR using specific forward and reverse primers followed by gel electrophoresis. Results Out of the total 720 samples, 38.9% (280/720) were culture positive. K. pneumoniae was the most predominant isolate 31.4% (88/280). Of 88 K. pneumoniae isolates, 56.8% (50/88) were multi-drug resistant (MDR), and 51.1% (45/88) were MHT positive. Colistin showed the highest sensitivity (100%; 88/88), followed by tigecycline (86.4%; 76/88). blaNDM-2 and blaOXA-48 genes were detected in 24.4% (11/45) and 15.5% (7/45) of carbapenemase-producing K. pneumoniae isolates, respectively. Conclusion The rate of MDR and carbapenemase production was high in the K. pneumoniae isolates. Colistin and tigecycline could be the drug of choice for the empirical treatments of MDR and carbapenemase-producing K. pneumoniae. Our study provides a better understanding of antibiotic resistance threat and enables physicians to select the most appropriate antibiotics.


2020 ◽  
Vol 25 (2) ◽  
pp. 49-54
Author(s):  
Rabin Gyawali ◽  
Ram Bahadur Khadka ◽  
Basudha Shrestha ◽  
Sarita Manandhar

Considerable increase in the prevalence and multidrug-resistant (MDR) Pseudomonas has been observed with towering morbidity and mortality. As a consequence of the haphazard use of antimicrobials, the spread of antimicrobial resistance is now a global issue. This study aimed to access the distribution rate and antibiotic susceptibility patterns of Pseudomonas species isolated from various clinical specimens in Kathmandu Model Hospital, Nepal. During the study period, 1252 samples were collected, cultured and the organism was isolated and identified. The antimicrobial susceptibility testing was done using the modified Kirby-Bauer disc diffusion method as per CLSI guidelines. Out of 1252 samples, 28 clinical isolates of Pseudomonas species were isolated. The highest number of Pseudomonas spp. was isolated from swab samples that included pus, ear, and wound (46.4 %). Pseudomonas spp. demonstrated marked resistance against cefixime (96.4 %) and showed higher sensitivity to piperacillin/tazobactam (92.9 %). The result showed pus, wound exudates, ear discharges samples exhibit Pseudomonas as common etiology of infection. Pseudomonas spp. demonstrated highest sensitivity against piperacillin/tazobactam, amikacin, meropenem, gentamycin. The steady resistance of Pseudomonas spp. to most of the antibiotics, necessitates these drugs to be confined to extreme infections and hospital intensive care units to circumvent the speedy emergence of resistant strains.


2007 ◽  
Vol 28 (8) ◽  
pp. 941-944 ◽  
Author(s):  
Thean Yen Tan ◽  
Karen Poh ◽  
Siew Yong Ng

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.


2010 ◽  
Vol 4 (12) ◽  
pp. 810-813 ◽  
Author(s):  
Fatima Kaleem ◽  
Javaid Usman ◽  
Afreenish Hassan ◽  
Aslam Khan

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.


Author(s):  
Mousumi Karmaker ◽  
Md. Abul Khair ◽  
Una Jessica Sarker ◽  
Rabeya Nahar Ferdous ◽  
Sa’dia Tasnim ◽  
...  

Pseudomonas aeruginosa is one of the most widespread gram-negative microorganisms identified in the clinical samples and most common causes of hospital acquired infection. P. aeruginosa is affecting both indoor and outdoor patients throughout the world. Due to frequent mutation in          P. aeruginosa highly resistant strain developed rapidly. The aim of the study to determine the prevalence of P. aeruginosa species in different samples isolated from a Tertiary care Hospital as well as determination their diverse antibiotic resistance pattern. This cross-sectional study was carried out to determine in-vitro resistance pattern of P. aeruginosa isolates to common antimicrobial agents by disc diffusion method. Various clinical samples were collected from Bangladesh Health Sciences Hospital (BIHS) General Hospital, Dhaka. This research was carried out in the Department of Microbiology of Bangladesh University of Health Sciences (BUHS). Isolation, identification and antibiogram were performed for P. aeruginosa following standard microbiological laboratory procedure. A total of 218 P. aeruginosa were isolated from 3062 different clinical specimens which are statistically significant (p<0.0001). Among the highest number of P. aeruginosa were isolated from outdoor patients 140 compare to Indoor patients which are significantly higher (p <0.013). In this study Male (68.3%) are more vulnerable to P. aeruginosa infection compare to females (31.7%) which is also statistically significant. Young people (less than 35 years) were more susceptible to P. aeruginosa infection which is also statistically significant (p< 0.01). The highest number of P. aeruginosa was isolated from wound (43.12%), followed by pus (40.33%), sputum (8.71%) urine (7.80%). The maximum number of P. aeruginosa in various samples was resistant to aztreonam and co-tromoxazole followed by cephalosporins, aminoglycosides, carbapenems. The most sensitive antibiotic was colistin of followed by gentamycin and tetracycline. To control the spread of resistant bacteria, it is disparagingly vital to have stringent antibiotic guidelines. The antibiotic susceptibility pattern of P. aeruginosa requires to be continuously monitored in specialized clinical units and the results readily made available to the clinicians to minimize the resistance.


2019 ◽  
Author(s):  
Wirittamulla Gamage Maheshika Kumudunie ◽  
Wijesooriya Rathnayaka Pathirennehalage Lakmini Inoka Wijesooriya ◽  
Kalubowilage Dhananja Namalie ◽  
Narapity Pathirannehalage Sunil-Chandra ◽  
Yasanandana Supunsiri Wijayasinghe

Abstract Background Extended spectrum β-lactamase producing Enterobacteriaceae (ESBL-PE) and carbapenamase producing Enterocacteriaceae (CPE) are widely disseminated globally creating a huge public health threat. Even though incidence of multidrug-resistant Enterobacteriaceae is rapidly growing, the epidemiological data regarding the occurrence of CRE in Sri Lanka is scarce. In this study, we determined the prevalence of ESBP-PE and CRE and the genetic determinants of CPE. Methods A total of 593 clinically significant Enterobacteriaceae was isolated from different clinical samples (urine, pus/wound, respiratory, blood, and other sterile specimens) at a tertiary care hospital in Sri Lanka from December 2017 – February 2018. Antimicrobial susceptibility and identification of ESBL-PE, CRE were done by disc diffusion method. CRE were identified to species level using a rapid identification kit and carbapenemase production was determined by modified carbapenem inactivation method. The presence of blaKPC, blaNDM, blaOXA-48-like genes were detected by PCR. Results The overall prevalence of ESBL-PE and CRE were found to be 26.0% and 9.6%, respectively. The rate of ESBL-PE in different sample types ranged from 18.2% to 30.8% with the highest prevalence among uropathogenic Enterobacteriaceae. The occurrence of CRE ranged from 6.7% to 20.8% with the highest prevalence among respiratory Enterobacteriaceae. CRE species identified were K. pneumoniae (80.7%), E. coli (5.3%), C. freundii (7.0%), P. rettgeri (3.5%), E. cloacae (1.7%), and E. aerogenes (1.7%). The carbapenemase production was detected in 94.7% of CRE isolates. The carbapenemases found were OXA-48-like (88.9%), NDM (14.8%), and KPC (3.7%). Conclusions The prevalence of CRE in Sri Lanka is alarming. Carbapenamse production was the major mechanism of carbapenem resistance and K. pneumoniae was the predominant CRE. Presence of KPC enzyme was detected in addition to the previously reported NDM and OXA-48-like carbapenamases in Sri Lanka. The rapid spread of CPE, necessitates the prompt implementation of preventive measures in Sri Lanka.


Author(s):  
Sulaiman A Mohammed ◽  
Dr. Suman P Singh

Background: Carbapenem resistance mediated by metallo beta lactamases (MBL) in Acinetobacter baumannii is a global challenge due to its rapid spread and limited therapeutic options. Objective: To determine the prevalence of MBL in A. baumannii isolates in hospitalized patients by both phenotypic and genotypic methods. Materials and Methods: The clinical samples were collected from inpatients and subcultured on routine culture media for growth. Identification of bacteria along with antimicrobial sensitivity testing was done by VITEK -2 Compact (bioMerieux). Antibiotics that were not tested by VITEK-2 were tested manually by Kirby-Bauer disk diffusion method according to CLSI 2017 and EUCAST 2016 guidelines. The isolates which were resistant to carbapenem (imipenem and/ or meropenem) were tested by phenotypic (imipenem-EDTA combined disk method) and genotypic method for presence of common metallo beta lactamases genes (blaIMP, blaNDM, blaGIM, blaVIM, blaSPM and blaSIM). Results: 84 non duplicate A.baumannii were isolated out of 947 pathogenic gram negative isolates. Majority (47.6%) of isolates were obtained from tracheostomy/endotracheal/bronchoalveolar lavage (TT/ET/BAL) followed by sputum (21.4%). None of the isolates were found to be resistant to colistin and tigecycline. 73 (86.9%) isolates were found to be carbapenem resistant, among these 60 (82.2%) were found to be MBL positive by phenotypic and 32 (43.2%) by genotypic method. MBL genes detected were blaNDM (39.7%), blaGIM (2.7%) and blaVIM (1.4%). None of the isolates were positive for blaIMP, blaSPM and blaSIM. Conclusion:  The prevalence of MBL in carbapenem resistant isolates of A.baumannii was 87.7%.  blaNDM was the most common gene detected. No significant difference was found in the ability of phenotypic and genotypic methods for MBL detection. The resistance rate of the A.baumannii is high for most antibiotics except for polymyxins (E&B) and tigecycline. Key words: Metallo beta Lactamases, Acinetobacter baumannii, Carbapenem.


2017 ◽  
Vol 3 (1) ◽  
pp. 11-15
Author(s):  
Sitesh Karn ◽  
Narayan Dutt Pant ◽  
Sanjeev Neupane ◽  
Saroj Khatiwada ◽  
Shaila Basnyat ◽  
...  

Background Carbapenems are considered as drugs of choice for the treatment of the infections caused by drug resistant bacteria. However, in the recent years the prevalence of carbapenem resistant gram negative bacteria has increased significantly. The main objective of this study was to determine the prevalence of carbapenemase producing gram negative bacteria among all the clinical isolates.Material and methods A total of 3246 non-repeated, different clinical specimens from patients attending Kathmandu Model Hospital, from July 2013 to January 2014 were cultured and the gram negative bacterial isolates obtained were subjected to identification with the help of colony morphology, Gram’s stain and conventional biochemical tests. Kirby-Bauer disk diffusion technique was used to perform antimicrobial susceptibility testing. Phenotypic confirmation of carbapenemase and AmpC beta-lactamase production was done by combined disc method.Results 890 samples showed the growth of bacterial pathogens. Out of total 769 gram negative bacteria, 57 were found to be carbapenem resistant. Of which, highest number (47) of the isolates were found to be metallo-β lactamase (MBL) producers. Six bacterial isolates produced both (Klebsiella pneumoniae carbapenemase) KPC and MBL, whereas only one isolate was found to be positive for both MBL and AmpC. Three bacterial strains showed carbapenem resistance due to over production of AmpC β-lactamase.Conclusion Among carbapenem resistant gram negative bacteria, MBL was present as the major enzyme responsible for resisting carbapenem antibiotics.


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