Detection of Klebsiella pneumoniae Carbapenemase (KPC) and Metallo-Beta Lactamase (MBL) Producing Gram Negative Bacteria Isolated from Different Clinical Samples in A Transplant Center, Kathmandu, Nepal (original) (raw)
Related papers
Indian Journal of Animal Research, Volume 57 Issue 4: 517-521 (April 2023)
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 Gramnegative 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-betalactamase (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.
IP Innovative Publication Pvt. Ltd., 2017
Introduction: Carbapenemases are one of the common β-lactamases seen in Klebsiella pneumoniae that are responsible for multi drug resistance. Detection of resistance in these bacteria is necessary for formulation of infection control policies. The hidden resistance of this bacteria is critical to diagnose. To address this problem, the present study aims to detect Carbapenemase production and antimicrobial susceptibility patterns in clinical isolates of Klebsiella pneumoniae at the present setting. Materials and Method: A total of 438 strains of Klebsiella pneumoniae were isolated and subjected to Carbapenemase detection by Screening method using Imepenem (10 µg) disc. Modified Hodge test and Combined disc diffusion test along with E-test was done to confirm carbapenemase production followed by antimicrobial susceptibility testing. E-test was considered to be gold standard. Results: Total 34/438 (7.76%) carbapenem resistant isolates were obtained. The carbapenemase positive isolates were predominantly isolated from Burns wards (14.61%). E-test considering it as gold standard test confirmed all 34 of these as carbapenemase producers. Out of 34, MHT detected 31and CDT detected 32 isolates as positive for carbapenemase production. They were highly resistant to cefotaxime, and ceftazidime (91.18%). Conclusion: Screening for Carbapenemase production needs to be carried out routinely in every clinical diagnostic facility. There is a need for rational use and strict adherence to the concept of " reserve drugs " to minimize the misuse of available antimicrobials. The findings of this study emphasize the need for a continuous surveillance in the ICUs and different hospital wards to detect the resistant strains.
Cureus, 2022
Background: Carbapenemase-producing Klebsiella pneumoniae (CRKP) has become a menace in several intensive care units, which needs to be controlled immediately after being reported by a laboratory. Detection in the laboratory is usually done using phenotypic methods and it is not known whether knowledge of these genes helps in individual patient management. This study aimed to compare the outcomes of oxacillinases β-lactamases (OXA-48) and New Delhi metallo-β-lactamase (NDM-1)-producing CRKP isolates, the two most common carbapenemases reported from India, obtained from patients with bloodstream infections in an ICU in a tertiary care center in North India and to compare the different laboratory methods for their detection. Materials and methods: Klebsiella pneumoniae isolates obtained from the blood culture of patients admitted to various ICUs were subjected to conventional polymerase chain reaction (PCRs) for blaNDM and blaOXA48-like genes. Those positive for any of the genes were tested by the modified carbapenem inactivation method (mCIM) and if found positive were also subjected to ethylenediamine tetraacetic acid (EDTA)-modified carbapenem inactivation method (eCIM). Antibiotic susceptibility tests (AST) were performed and clinical data were recorded. Results: A total of 49 isolates were positive for one or more carbapenemase genes (30 {61.2%} for blaNDM gene only, 13 {26.5%)} for blaOXA48-like gene only, and six {12.2%} for both). The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of mCIM were found to be 77.6%, 100%, 100%, and 78.9%, respectively. Statistically significant differences were found in the AST pattern between the isolates with two genes. Increased MIC levels of colistin were observed, though they lay in the sensitive range. Mortality occurred in all six patients who were infected with CRKP harboring both the genes though no significant difference was observed in NDM and OXA-48 producing CRKP isolates. Conclusion: Surveillance of carbapenemase genes in a hospital setting is essential. The possible reasons for the low diagnostic accuracy of mCIM and differences in AST patterns are discussed.
Journal of Antimicrobial Chemotherapy, 2017
We evaluated the in vitro activity of different antimicrobial combinations with and without colistin against 39 carbapenem-resistant Klebsiella pneumoniae (CR-Kp) strains (colistin ! meropenem/ doripenem, colistin ! tigecycline, colistin ! rifampicin, gentamicin ! meropenem, gentamicin ! tigecycline and the double-carbapenem regimen meropenem ! ertapenem) using the chequerboard method. The triple combination colistin ! meropenem ! tigecycline was also tested. In addition, killing studies were performed for meropenem ! ertapenem. Results: Gentamicin-based combinations showed a high level of synergy. Meropenem ! ertapenem was synergic in 12/39 (30.7%) of the strains, whereas based on killing studies 1 % MIC meropenem ! 1 % MIC ertapenem and 2 % MIC meropenem ! 1 % MIC ertapenem combinations were bactericidal and synergic at 24 h [mean area under the bactericidal curve (AUBC) 54.9+26.1 and 44.2+15.3 compared with 1 % MIC meropenem (134.5+40.1) and 2 % MIC meropenem (126.4+5.4), respectively, P , 0.0001]. When the results were stratified according to meropenem MIC, we found that the degree of synergy significantly increased for isolates with lower meropenem (and not ertapenem) MICs, up to an MIC of 128 mg/L. Among colistin-containing combinations, synergy was observed in 18/39 (46.1%), 33/34 (97%), 24/39 (61.5%) and 17/39 (43.5%) of the strains for colistin ! meropenem, colistin ! rifampicin, colistin ! tigecycline and colistin ! doripenem, respectively, including colistinresistant strains. Colistin ! meropenem ! tigecycline at subinhibitory concentrations resulted in the absence of growth of 37/39 strains (94.8%). Conclusions: Our in vitro data suggest that colistin might be a valid therapeutic option against CR-Kp, even in the presence of colistin resistance, whereas the double-carbapenem regimen represents a viable option when colistin is not recommended, especially if the meropenem MIC is 128 mg/L. Since traditional antimicrobial susceptibility reports are not sufficiently informative for clinicians, synergy testing as well as actual meropenem MIC evaluation should always be performed in the case of CR-Kp infections.
Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]
The production of KPC (Klebsiella pneumoniae carbapenemase) is the major mechanism of resistance to carbapenem agents in enterobacterias. In this context, forty KPC-producing Enterobacter spp. clinical isolates were studied. It was evaluated the activity of antimicrobial agents: polymyxin B, tigecycline, ertapenem, imipenem and meropenem, and was performed a comparison of the methodologies used to determine the susceptibility: broth microdilution, Etest(®) (bioMérieux), Vitek 2(®) automated system (bioMérieux) and disc diffusion. It was calculated the minimum inhibitory concentration (MIC) for each antimicrobial and polymyxin B showed the lowest concentrations for broth microdilution. Errors also were calculated among the techniques, tigecycline and ertapenem were the antibiotics with the largest and the lower number of discrepancies, respectively. Moreover, Vitek 2(®) automated system was the method most similar compared to the broth microdilution. Therefore, is important to evalua...
Treatment options are limited in infections caused by extended-spectrum -lactamase (ESBL)-producing Enterobacteriaceae, with carbapenems generally preferred. Disturbingly, however, carbapenem-resistant strains are emerging worldwide. Here we report two clinical isolates, one Escherichia coli and one Klebsiella pneumoniae, each with high-level carbapenem resistance (imipenem minimum inhibitory concentration of 32 g/mL). They were isolated following imipenem therapy from two hospital patients who had received imipenem therapy in different regions of Turkey. Both isolates produced OXA-48-like carbapenemases, enzymes so far reported only from Turkey. Both isolates also had group 1 CTX-M-type ESBLs and had lost major outer membrane proteins. OXA-48-like carbapenemases appear to be scattered in Turkey and surveillance to determine their prevalence is warranted.
Hippokratia
New Delhi metallo-beta-lactamase-1(NDM-1) is a novel type of metallo-beta-lactamase (MBL) which inactivates all β-lactam antibiotics except aztreonam. Enterobacteriaceae expressing NDM-1 have been identified worldwide. The aim of this study was to detect MBLs in carbapenem-resistant K. pneumoniae isolates obtained from patients hospitalized in one of the university hospitals in Isfahan, Iran. Of the 112 isolates obtained from various clinical samples, 49 were selected for carbapenemase detection based on their reduced susceptibility to imipenem or meropenem according to the disc diffusion method. These isolates were screened for carbapenemase and MBL production using the Modified Hodge Test (MHT) and Epsilometer test (E-test) MBL strips. Polymerase chain reaction was performed on all 49 isolates using specific primers to detect genes encoding IMP (active on imipenem), VIM (Verona integron-encoded metallo-β-lactamase), SPM-1 (Sao Paulo metallo-β-lactamase) and NDM-1. Among 49 carbapen...
Antimicrobial Agents and Chemotherapy, 1999
An important mechanism of bacterial resistance to -lactam antibiotics is inactivation by -lactam-hydrolyzing enzymes (-lactamases). The evolution of the extended-spectrum -lactamases (ESBLs) is associated with extensive use of -lactam antibiotics, particularly cephalosporins, and is a serious threat to therapeutic efficacy. ESBLs and broad-spectrum -lactamases (BDSBLs) are plasmid-mediated class A enzymes produced by gram-negative pathogens, principally Escherichia coli and Klebsiella pneumoniae. MK-0826 was highly potent against all ESBL-and BDSBL-producing K. pneumoniae and E. coli clinical isolates tested (MIC range, 0.008 to 0.12 g/ml). In E. coli, this activity was associated with high-affinity binding to penicillin-binding proteins 2 and 3. When the inoculum level was increased 10-fold, increasing the amount of -lactamase present, the MK-0826 MIC range increased to 0.008 to 1 g/ml. By comparison, similar observations were made with meropenem while imipenem MICs were usually less affected. Not surprisingly, MIC increases with noncarbapenem -lactams were generally substantially greater, resulting in resistance in many cases. E. coli strains that produce chromosomal (Bush group 1) -lactamase served as controls. All three carbapenems were subject to an inoculum effect with the majority of the BDSBL-and ESBL-producers but not the Bush group 1 strains, implying some effect of the plasmid-borne enzymes on potency. Importantly, MK-0826 MICs remained at or below 1 g/ml under all test conditions.
A Study on Carbapenemase Producing Klebsiella Pneumoniae In Beni Suef University Hospital
International Journal of Scientific Research in Science and Technology, 2022
Background: Widespread dissemination of Klebsiella pneumoniae carbapenemase (KPC) is of major concern in healthcare settings. Resistance to carbapenems involves multiple mechanisms such as the production of carbapenemases, impermeability of outer membrane and efflux pump mechanism. Objective: The aim of this study was to evaluate the prevalence of carbapenemase-producing K. pneumoniae strains among various clinical specimens obtained from different wards and to detect KPC as a mechanism of resistance. Methods: 100 samples of urine and sputum (55 urine and 45sputum) were collected from outpatients and inpatients attending urology and chest departments in Beni Suef University Hospital aiming to isolate K.pneumniae during the period of December 2016 through January 2018. The isolates were tested for susceptability to ertapenem using E test. Resistant isolates were subjected to phenotypic detection of carbapenemase production by MHT and molecular assessment of KPC gene by PCR. Phylogentic tree was used to detect their relationship. Results: K.pneumonia were isolated from 31(31%) of the samples taken. Out of them 19(61.8%) were resistant to ertapenem. By MHT,17/19 (89.4%) were positive for carbapenemase; and only 13 out of them (76.4%) were confirmed as KPC by PCR. Conclusion: High rate of carbapenem- resistance in K. pneumoniae by both phenotypic and molecular methods. Initiating appropriate infection control measures along with a strictly implemented antibiotic stewardship program are necessary to prevent their spread.
2013
Treatment options are limited in infections caused by extended-spectrum -lactamase (ESBL)-producing Enterobacteriaceae, with carbapenems generally preferred. Disturbingly, however, carbapenem-resistant strains are emerging worldwide. Here we report two clinical isolates, one Escherichia coli and one Klebsiella pneumoniae, each with high-level carbapenem resistance (imipenem minimum inhibitory concentration of 32 g/mL). They were isolated following imipenem therapy from two hospital patients who had received imipenem therapy in different regions of Turkey. Both isolates produced OXA-48-like carbapenemases, enzymes so far reported only from Turkey. Both isolates also had group 1 CTX-M-type ESBLs and had lost major outer membrane proteins. OXA-48-like carbapenemases appear to be scattered in Turkey and surveillance to determine their prevalence is warranted.