scholarly journals Co occurrence of two 16S rRNA methyltrasferases along with NDM and OXA 48 like carbapenamases on a single plasmid in Klebsiella pneumoniae

2019 ◽  
Vol 11 (04) ◽  
pp. 305-311 ◽  
Author(s):  
K. V. L. Aishwarya ◽  
P. V. Geetha ◽  
M. Shanthi ◽  
S. Uma
Keyword(s):  
16S Rrna ◽  
Klebsiella Pneumoniae ◽  
Conjugative Plasmid ◽  
Beta Lactams ◽  
Plasmid Replicon ◽  
Single Plasmid ◽  
Group A ◽  
Polymerase Chain ◽  
Encoding Genes ◽  
Conjugation Experiment

Abstract BACKGROUND: The carbapenemase-encoding genes, bla NDM- and bla OXA-48 - like , confer resistance to all the known beta-lactams and are encountered along with other beta-lactamase-encoding genes and/or 16S ribosomal RNA (rRNA)-methylating genes. The co-occurrence of bla NDM and bla OXA-48 - like on a single plasmid is a rare occurrence. AIM AND OBJECTIVE: The purpose of the study was to characterize the plasmids in Klebsiella pneumoniae isolates producing 16S rRNA methyltransferase along with bla NDM , bla OXA-48-like , and other resistance encoding genes. MATERIALS AND METHODS: One-hundred and seventeen K. pneumoniae clinical isolates which were resistant to aminoglycosides were collected. Polymerase chain reaction-based screening for 16S rRNA methyltransferase genes armA, rmtB, and rmtC; carbapenamase genes bla NDM , bla OXA-48-like , bla IMP, bla VIM, and bla KPC ; and other resistance genes such as bla TEM, bla SHV, bla CTX-M , and qnr (A, B, and S) determinants acc (6') Ib-cr was performed. Conjugation experiment was carried out for seven isolates that anchored bla NDM and bla OXA-48-like along with any one of the 16S rRNA methyltransferases. The plasmid-based replicon typing for different plasmid-incompatible (Inc) group was performed on the conjugatively transferable plasmids. RESULTS: Among the 16S rRNA methyltransferases, armA was more predominant. bla NDM and bla OXA-48 -like were present in 56 (47.86%) and 22 (18.80%) isolates, respectively. Out of seven isolates which were conjugatively transferable, only four had bla NDM and bla OXA-48 - like on the same plasmid and they belonged to Inc N and A/C replicon. Three isolates co-harbored 16S rRNA methyltransferases armA, rmtB, and rmtC, and out of the them, one isolate harbored two 16S rRNA methyltransferases armA and rmtB, on the single-plasmid replicon A/C. CONCLUSION: This is the first report revealing the coexistence of bla NDM and bla OXA-48 - like co-harboring two 16S rRNA methylases on a single conjugative plasmid replicon belonging to incompatibility group A/C.

scholarly journals Modified CIM-Test As a Useful Tool to Detect Carbapenemase Activity Among Extensively Drug- Resistant Klebsiella Pneumoniae, Escherichia Coli and Acinetobacter Baumannii

2021 ◽  
Author(s):  
Abed Zahedi bialvaei ◽  
Alireza Dolatyar Dehkharghani ◽  
Farhad Asgari ◽  
Firouzeh Shamloo ◽  
Parisa Eslami ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Acinetobacter Baumannii ◽  
Sensitivity And Specificity ◽  
E Coli ◽  
Carbapenem Resistant ◽  
Polymerase Chain ◽  
Phenotypic Methods ◽  
Encoding Genes ◽  
Positive Results

Abstract Background Timely detection of carbapenemases is essential for developing strategies to control the spread of infections by carbapenem-resistant isolates. The purpose of our study was to determine the epidemiology of carbapenemase genes among carbapenem resistant isolates of Acinetobacter baumannii, Klebsiella pneumoniae and Escherichia coli and to compare efficacy of modified Hodge Test (MHT), Carba NP and modified carbapenem inactivation method (mCIM) tests. Methods A total of 122 carbapenem-resistant clinical isolates including 77 K. pneumoniae, 39 A. baumannii, and six E. coli were collected from hospitalized patients. Three phenotypic methods, including MHT, Carba NP test and mCIM were used for investigation of carbapenemase production. In addition, polymerase chain reaction (PCR) was performed to detect carbapenemase encoding genes. Results The sensitivity and specificity of the MHT were 75.0% and 100% respectively. In addition, CarbaNP displayed 80.8% sensitivity and 100% specificity, whereas the sensitivity and specificity were 90.4% and 100% for the mCIM test, respectively. Among carbapenem-resistant isolates, 70, 84 and 87 isolates exhibited positive results according to MHT, CarbaNP test and mCIM, respectively. PCR indicated the presence of one or more carbapenemase genes in 119 of carbapenem-resistant isolates, with blaKPC and blaVIM being the most commonly encountered. Co-production of ‘KPC and VIM’, ‘KPC and IMP’ and ‘KPC and OXA-48’ was detected in nine, seven and three isolates, respectively. Conclusion Our results confirm that the mCIM test is a useful tool for the reliable detection of carbapenemases-activity in enterobacterial isolates, especially in clinical microbiological laboratories with limited resources.

scholarly journals Modified CIM test as a useful tool to detect carbapenemase activity among extensively drug-resistant Klebsiella pneumoniae, Escherichia coli and Acinetobacter baumannii

2021 ◽  
Vol 71 (1) ◽  
Author(s):  
Abed Zahedi Bialvaei ◽  
Alireza Dolatyar Dehkharghani ◽  
Farhad Asgari ◽  
Firouzeh Shamloo ◽  
Parisa Eslami ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Acinetobacter Baumannii ◽  
Sensitivity And Specificity ◽  
E Coli ◽  
Carbapenem Resistant ◽  
Polymerase Chain ◽  
Phenotypic Methods ◽  
Encoding Genes ◽  
Positive Results

Abstract Purpose Timely detection of carbapenemases is essential for developing strategies to control the spread of infections by carbapenem-resistant isolates. The purpose of this study was to determine the epidemiology of carbapenemase genes among carbapenem-resistant isolates of Acinetobacter baumannii, Klebsiella pneumoniae, and Escherichia coli. In addition, the efficacy of the modified Hodge test (MHT), Carba NP test, and modified carbapenem inactivation method (mCIM) were compared. Methods A total of 122 carbapenem-resistant clinical isolates including 77 K. pneumoniae, 39 A. baumannii, and six E. coli were collected from hospitalized patients. Three phenotypic methods, including the MHT, Carba NP test, and mCIM were used for investigation of carbapenemase production. In addition, polymerase chain reaction (PCR) was performed to detect carbapenemase-encoding genes. Result The sensitivity and specificity of the MHT were 75.0% and 100%, respectively. In addition, Carba NP displayed 80.8% sensitivity and 100% specificity, whereas the sensitivity and specificity were 90.4% and 100% for the mCIM test, respectively. Among carbapenem-resistant isolates, 70, 84, and 87 isolates exhibited positive results according to the MHT, Carba NP test, and mCIM, respectively. PCR indicated the presence of one or more carbapenemase genes in 119 of carbapenem-resistant isolates, with blaKPC and blaVIM being the most commonly encountered. Co-production of ‘KPC and OXA-48’, ‘KPC and VIM’, and ‘KPC and IMP’ was detected in three, nine, and seven isolates, respectively. Conclusion Our results confirm that the mCIM test is a useful tool for the reliable detection of carbapenemase activity in enterobacterial isolates, especially in clinical microbiological laboratories with limited resources.

scholarly journals Prevalence of Transferable OXA-1 β-Lactamase Associated with Carbapenem-Resistant Klebsiella pneumoniae Isolates in Iraq

2021 ◽  
Author(s):  
Fatima Moeen Abbas
Keyword(s):  
Klebsiella Pneumoniae ◽  
Diffusion Method ◽  
Antibiotics Resistance ◽  
Disk Diffusion Method ◽  
Antibiotic Resistance Profile ◽  
Carbapenem Resistant ◽  
Stool Samples ◽  
Polymerase Chain ◽  
High Level ◽  
Conjugation Experiment

This study was designed to explore the incidence of blaOXA-1 amongst Klebsiella pneumoniae isolates with resistant to carbapenem. Between December 2014 and April 2015, one hundred samples were taken from two hospitals: Babylon Teaching Hospital for Maternity and Pediatric / Babylon Province (clinical, umbilical infections, n= 40; environmental, n=20) and Karbala Hospital for Pediatric / Karbala Province (40 stool samples). All patients were hospitalized or attended these hospitals, all under 1 year of age. Seventeenth (17%) isolates were identified as Klebsiella pneumoniae. The antibiotic resistance profile of isolates was tested using disk diffusion method. High-level of resistance was recorded with ampicillin (94.1%) and piperacillin (88.2%) antibiotics. Resistance to carbapenem was reported in two K.pneumoniae isolates, these were investigated for the existence of OXA-1b-lactamase using Polymerase Chain Reaction (PCR) technique. Two (100%) isolates gave positive result. Transference of this gene was studied by conjugation experiment. The blaOXA-1 gene conjugated successfully in 1 (50%) isolate only.

scholarly journals Detection of 16S rRNA Methylases and Co-Resistance with β-lactams among Klebsiella pneumoniae Isolates from Iraqi Patients

2019 ◽  
Vol 16 (3) ◽  
pp. 0580
Author(s):  
Mustafa Et al.
Keyword(s):  
Amino Acids ◽  
16S Rrna ◽  
Klebsiella Pneumoniae ◽  
Biochemical Properties ◽  
Clinical Samples ◽  
Missense Mutations ◽  
Accession Number ◽  
Beta Lactamases ◽  
Extended Spectrum Beta Lactamases ◽  
Encoding Genes

Out of 150 clinical samples, 50 isolates of Klebsiella pneumoniae were identified according to morphological and biochemical properties. These isolates were collected from different clinical samples, including 15 (30%) urine, 12 (24%) blood, 9 (18%) sputum, 9 (18%) wound, and 5 (10%) burn. The minimum inhibitory concentrations (MICs) assay revealed that 25 (50%) of isolates were resistant to gentamicin (≥16µg/ml), 22 (44%) of isolates were resistant to amikacin (≥64 µg/ml), 21 (42%) of isolates were resistant to ertapenem (≥8 µg/ml), 18 (36%) of isolates were resistant to imipenem (4- ≥16µg/ml), 43 (86%) of isolates were resistant to ceftriaxone (4- ≥64 µg/ml), 42 (84%) of isolates were resistant to ceftazidime (16-64 µg/ml), and 40 (80%) of isolates were resistant to cefepime (4- ≥16µg/ml). Co-Resistance for both β-lactams and aminoglycosides were detected among 25 (50%) of K. pneumoniae isolates. The extended spectrum beta-lactamases (ESBLs) were detected among 25 (50%) of K. pneumoniae isolates. Screening of 16S rRNA methylases  encoding genes revealed that armA was found in 5 (10%) of K. pneumoniae isolates, whereas rmtB was not found among K. pneumoniae isolates. DNA sequencing of armA revealed that the presence of missense mutations in which affected in the translation of protein by substitutions of amino acids, leading to increase the resistance values of MICs for gentamicin and amikacin. These variants were registered in NCBI at the accession number LC373258. The phylogenetic tree of armA variants showed a slight deviation of these variants from K. pneumoniae species.

Polymerase chain reaction amplification and typing of rotavirus in environmental samples

1995 ◽  
Vol 31 (5-6) ◽  
pp. 371-374 ◽  
Author(s):  
R. Gajardo ◽  
R. M. Pintó ◽  
A. Bosch
Keyword(s):  
Polymerase Chain Reaction ◽  
Environmental Samples ◽  
Polymerase Chain Reaction Amplification ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Group A ◽  
Reaction Amplification ◽  
Stool Specimens ◽  
Polymerase Chain

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.

scholarly journals Wastewaters, with or without Hospital Contribution, Harbour MDR, Carbapenemase-Producing, but Not Hypervirulent Klebsiella pneumoniae

Antibiotics ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 361
Author(s):  
Adela Teban-Man ◽  
Anca Farkas ◽  
Andreea Baricz ◽  
Adriana Hegedus ◽  
Edina Szekeres ◽  
...  
Keyword(s):  
Principal Component Analysis ◽  
Klebsiella Pneumoniae ◽  
Wastewater Treatment Plants ◽  
Principal Component ◽  
Drug Resistant ◽  
Beta Lactams ◽  
Carbapenemase Gene ◽  
Antibiotic Susceptibility Patterns ◽  
Sequence Types ◽  
Susceptibility Patterns

Carbapenemase-producing Klebsiella pneumoniae (CPKP) isolated from influent (I) and effluent (E) of two wastewater treatment plants, with (S1) or without (S2) hospital contribution, were investigated. The strains belonged to the Kp1 phylogroup, their highest frequency being observed in S1, followed by S2. The phenotypic and genotypic hypervirulence tests were negative for all the strains tested. At least one carbapenemase gene (CRG), belonging to the blaKPC, blaOXA-48, blaNDM and blaVIM families, was observed in 63% of CPKP, and more than half co-harboured two to four CRGs, in different combinations. Only five CRG variants were observed, regardless of wastewater type: blaKPC-2, blaNDM-1, blaNDM-6, blaVIM-2, and blaOXA-48. Sequence types ST258, ST101 and ST744 were common for both S1 and S2, while ST147, ST525 and ST2502 were found only in S1 and ST418 only in S2. The strains tested were multi-drug resistant (MDR), all being resistant to beta-lactams, cephalosporins, carbapenems, monobactams and fluoroquinolones, followed by various resistance profiles to aminoglycosides, trimethoprim-sulphamethoxazole, tigecycline, chloramphenicol and tetracycline. After principal component analysis, the isolates in S1 and S2 groups did not cluster independently, confirming that the antibiotic susceptibility patterns and gene-type profiles were both similar in the K. pneumoniae investigated, regardless of hospital contribution to the wastewater type.

scholarly journals Comparison of Three Rapid Commercial Canine Parvovirus Antigen Detection Tests with Electron Microscopy and Polymerase Chain Reaction

2009 ◽  
Vol 21 (3) ◽  
pp. 344-345 ◽  
Author(s):  
Silke Schmitz ◽  
Christina Coenen ◽  
König Matthias ◽  
Thiel Heinz-Jürgen ◽  
Reto Neiger
Keyword(s):  
Electron Microscopy ◽  
Polymerase Chain Reaction ◽  
Chronic Diarrhea ◽  
Gastrointestinal Disease ◽  
Clinical Signs ◽  
Canine Parvovirus ◽  
Chain Reaction ◽  
Group A ◽  
Polymerase Chain ◽  
Group B

Different antibody-based tests for rapid detection of Canine parvovirus antigens in feces are commercially available, allowing quick diagnosis in a clinical setting. However, the diagnostic accuracy of these tests compared with standard methods has not been evaluated so far. In the current study, 3 commercial tests were compared with immune-electron microscopy (IEM) and polymerase chain reaction (PCR). Dogs were divided into 3 groups: group A, samples from dogs with acute hemorrhagic diarrhea ( n = 50); group B, dogs with chronic diarrhea ( n = 10); and group C, dogs with no evidence of gastrointestinal disease ( n = 40). Specificity of all 3 commercial tests versus PCR and IEM was good to excellent (92.2–100%). Sensitivity, in contrast, was poor: 15.8–26.3% versus PCR and 50–60% versus IEM. In group A, 10 dogs were positive by IEM and 24 dogs were positive by PCR. Positive PCR results were also obtained from animals in control groups (group B, 1 dog; group C, 5 dogs). No dog in group B or C was positive by IEM. In conclusion, the rapid tests are useful to diagnose canine parvoviral enteritis, but they do not rule out parvovirus infection in an animal with typical clinical signs. In addition, a small percentage of healthy dogs and dogs with chronic diarrhea showed positive PCR results; this may be due to asymptomatic/persistent infection or intestinal passage of virus. The significance of this finding remains unclear.

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