scholarly journals Molecular Typing of Klebsiella pneumoniae Clinical Isolates by Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction

2020 ◽  
Vol 2020 ◽  
pp. 1-5
Author(s):  
Parinaz Sedighi ◽  
Omid Zarei ◽  
Kiana Karimi ◽  
Mohammad Taheri ◽  
Pezhman Karami ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Polymerase Chain Reaction ◽  
Klebsiella Pneumoniae ◽  
Molecular Typing ◽  
Genetic Relatedness ◽  
Clinical Isolates ◽  
Enterobacterial Repetitive Intergenic Consensus ◽  
Chain Reaction ◽  
Microbiological Methods ◽  
Polymerase Chain

Aim. Klebsiella pneumoniae is one of the most important causes of nosocomial infections, including pneumonia, sepsis, and urinary tract infection. Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) technique is a quick, reliable, and cost-effective method for molecular typing of Enterobacteriaceae family members. This study aimed to detect genetic relatedness among K. pneumoniae isolates from hospitals in Hamadan city, using ERIC-PCR technique. Materials and Methods. A total of 72 K. pneumoniae isolates were collected from patients admitted to Besat and Sina hospitals. After detection and confirmation of K. pneumonia isolates by chemical and conventional microbiological methods, DNAs were extracted after 24 hours of incubation at 37°C, using the boiling method. ERIC-PCR technique was carried out, and the ERIC patterns were analyzed by online data analysis service (inslico.ehu.es). ERIC profiles were compared using Dice method and clustered by UPGMA (unweighted pair group method with arithmetic mean) program. Also, the samples were evaluated by PCR method for the detection of aerobactin gene within their genome. Finding. The genetic relatedness among K. pneumoniae isolates was studied, and results established the genetic diversity of the clinical isolates by detecting 25 different ERIC types, including 14 common types and 11 unique types. Also, none of the isolates had aerobactin gene. Discussion. The results of this study showed high genetic diversity among K. pneumoniae strains, indicating the polyclonal distribution of K. pneumoniae isolates in Hamadan hospitals. This diversity causes problems for the treatment of infections due to the circulation of diverse K. pneumoniae clones, which possibly have different antimicrobial susceptibility patterns.

scholarly journals The Inc FII Plasmid and its Contribution in the Transmission of blaNDM-1 and blaKPC-2 in Klebsiella pneumoniae in Egypt

Antibiotics ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 266 ◽  
Author(s):  
Eman Ramadan Mohamed ◽  
Mamdouh Yones Ali ◽  
Nancy G F M Waly ◽  
Hamada Mohamed Halby ◽  
Rehab Mahmoud Abd El-Baky
Keyword(s):  
Polymerase Chain Reaction ◽  
Klebsiella Pneumoniae ◽  
Molecular Typing ◽  
University Hospital ◽  
Chain Reaction ◽  
Great Problem ◽  
E Coli ◽  
String Test ◽  
Resistance Patterns ◽  
Polymerase Chain

The emergence of blaKPC-2 and blaNDM-1 producing Klebsiella pneumoniae represents a great problem in many Egyptian hospitals. One hundred and twenty-six K. pneumoniae isolates from patients admitted to Assiut University Hospital were identified by an API20E kit. Carbapenemase-producing K. pneumoniae (CPKP) was detected by the modified carbapenem inactivation method (mCIM), the EDTA-modified carbapenem inactivation method (eCIM), and an E-test. Based on the polymerase chain reaction, all isolates were negative for bla-VIM-1 and bla-IMP-1, fifteen of these isolates were positive for both blaKPC-2 and blaNDM-1, two isolates were positive for blaKPC-2 only, and twenty-eight isolates were positive for bla-NDM-1 only. Although one isolate was positive for the string test, all CPKP isolates were negative for capsular genes. Only 71.1% of CPKP transferred their plasmids to their corresponding transconjugants (E. coli J53). The resistance patterns of the clinical isolates and their transconjugates were similar, except for 12 isolates, which showed differences with their transconjugates in the resistance profile of four antibiotics. Molecular typing of the plasmids based on replicon typing showed that Inc FIIK and FII plasmids predominated in isolates and their transconjugants carrying blaKPC-2 and/or blaNDM-1. Conjugative Inc FII plasmids play an important role in the spread of CPKP, and their recognition is essential to limit their spread.

Molecular typing of Acinetobacter baumannii clinical strains by enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR)

Gene Reports ◽  
2020 ◽  
Vol 18 ◽  
pp. 100542
Author(s):  
Elham Zarifi ◽  
Mehran Ghazalibina ◽  
Shamsoddin Mansouri ◽  
Korosh Morshedi ◽  
Roya Pourmajed ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Acinetobacter Baumannii ◽  
Molecular Typing ◽  
Enterobacterial Repetitive Intergenic Consensus ◽  
Chain Reaction ◽  
Clinical Strains ◽  
Polymerase Chain

scholarly journals Genetic Characterization Among Carbapenem-resistant Escherichia coli Strains Using Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (ERIC-PCR) Fingerprinting in South Africa

2020 ◽  
Author(s):  
Kingsley Ehi Ebomah ◽  
Anthony Ifeanyi Okoh
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Antimicrobial Agents ◽  
Genetic Relatedness ◽  
Carbapenem Resistance ◽  
Beta Lactam ◽  
Enterobacterial Repetitive Intergenic Consensus ◽  
Chain Reaction ◽  
E Coli ◽  
Polymerase Chain

Abstract Background Carbapenems belong to beta-lactam class of antibiotics usually considered as the last line of defense because they can be effective against severe infections caused by prevalent multidrug-resistant (MDR) pathogens. However, carbapenems can be deactivated by bacteria that produce carbapenemase (beta-lactamase). This study was conducted to screen for carbapenem-resistance genes (CRGs) harbored by pathogenic strains of Escherichia coli recovered from different environmental samples. We also assessed the genetic relatedness among selected E. coli pathotypes using enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR).Method: Molecular identification and characterization of the presumptive isolates were performed using PCR and isolates that exhibited antimicrobial resistance (AMR) phenotypically were further screened for some relevant CRGs (blaNDM−1, blaKPC and blaOXA−48−like). Furthermore, ERIC-PCR was used to determine the similarity and diversity of 31 E. coli strains which were randomly selected from the different sources analyzed in this study.Result Our findings revealed a total of 238 presumptive E. coli isolates, out of which 192 were confirmed positive for uidA gene. Further screening revealed 77 (40%) isolates belong to six key E. coli pathotypes and 70 of them exhibited phenotypic AMR. Additionally, twenty-nine (41%) of the 70 MDR pathogenic E. coli strains harbored CRGs; with 24 strains harboring blaNDM−1, 8 harboring blaKPC and 2 harboring blaOXA−48−like genes.Conclusion Findings also suggest that the selected E. coli pathotypes belonged to different genomic clusters, while the cluster analysis showed a possible genetic diversity among aquatic and farm isolates. Proper treatment of final effluents before discharge as well as the development of more effective strategies to control and manage the use of antimicrobial agents were strongly recommended.

scholarly journals Genetic Diversity of Klebsiella spp. Isolated from Tempe based on Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction (ERIC-PCR)

2013 ◽  
Vol 20 (4) ◽  
pp. 171-176 ◽  
Author(s):  
TATI BARUS ◽  
IVAN HANJAYA ◽  
JOANITA SADELI ◽  
BIBIANA WIDIYATI LAY ◽  
ANTONIUS SUWANTO ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Polymerase Chain Reaction ◽  
Enterobacterial Repetitive Intergenic Consensus ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Klebsiella Spp

Genetic Analysis ofPseudomonas aeruginosaby Enterobacterial Repetitive Intergenic Consensus Polymerase Chain Reaction (PCR) and Arbitrarily Primed PCR: Gel Analysis Compared with Microchip Gel Electrophoresis

2004 ◽  
Vol 25 (1) ◽  
pp. 65-71 ◽  
Author(s):  
Roudabeh J. Jamasbi ◽  
Stephen J. Kennel ◽  
Larry C. Waters ◽  
Linda J. Foote ◽  
J. Michael Ramsey
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetic Analysis ◽  
Gel Electrophoresis ◽  
Dna Analysis ◽  
Clinical Isolates ◽  
Enterobacterial Repetitive Intergenic Consensus ◽  
Chain Reaction ◽  
Electrophoretic Patterns ◽  
Pcr Products ◽  
Polymerase Chain

AbstractObjectives:To assess the applicability of a newly emerging microchip gel electrophoresis for rapid strain differentiation among clinical isolates ofPseudomonas aeruginosa,and to compare this technique with the traditional gel method for DNA separation.Methods:One hundred clinical strains ofP. aeruginosaobtained from a hospital in northwestern Ohio were tested for reactivity to 3 serotype-specific monoclonal antibodies by enzyme-linked immunosorbent assay. Twelve strains (4 from each serogroup) were selected for DNA analysis by polymerase chain reaction (PCR)-based, single primer DNA fingerprinting methods with 3 different primers: 1 enterobacterial repetitive intergenic consensus PCR and 2 arbitrarily primed PCRs. The PCR products were analyzed by agarose slab gel and microchip gel electrophoresis.Results:Of the 100 clinical isolates tested, 39% (4%, 14%, and 21%) were found to be serotypes 0:3, 0:6, and 0:11, respectively. Twelve strains were chosen for DNA analysis by PCR. The PCR products were analyzed by agarose slab gel electrophoresis and on microchips to determine interspecies diversity. Both methods demonstrated that different serotypes exhibited different electrophoretic patterns. Two strains (clinical strains 6 and 7, serotype 0:6) showed identical patterns, indicating a high degree of relatedness.Conclusion:In all cases, there was concordance between the electrophoretic patterns detected by the two methods. The capability of conducting both PCR and microchip gel electrophoresis offers an opportunity for an automated and rapid method for genetic analysis and differentiation among strains ofP. aeruginosaand other microorganisms.

scholarly journals Carbapenemase Production and Epidemiological Characteristics of Carbapenem-Resistant Klebsiella pneumoniae in Western Chongqing, China

2022 ◽  
Vol 11 ◽  
Author(s):  
Wan Huang ◽  
Jisheng Zhang ◽  
Lingyi Zeng ◽  
Chengru Yang ◽  
Lining Yin ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Klebsiella Pneumoniae ◽  
Virulence Genes ◽  
Resistance Rate ◽  
Molecular Characteristics ◽  
Chain Reaction ◽  
Detection Rates ◽  
Carbapenem Resistant ◽  
Polymerase Chain ◽  
Epidemiological Characteristics

BackgroundThis study aimed to determine the molecular characteristics of carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates in a hospital in western Chongqing, southwestern China.MethodsA total of 127 unique CRKP isolates were collected from the Yongchuan Hospital of Chongqing Medical University, identified using a VITEK-2 compact system, and subjected to microbroth dilution to determine the minimal inhibitory concentration. Enterobacteriaceae intergenic repeat consensus polymerase chain reaction and multilocus sequence typing were used to analyze the homology among the isolates. Genetic information, including resistance and virulence genes, was assessed using polymerase chain reaction. The genomic features of the CRKP carrying gene blaKPC-2 were detected using whole-genome sequencing.ResultsST11 was the dominant sequence type in the homology comparison. The resistance rate to ceftazidime-avibactam in children was much higher than that in adults as was the detection rate of the resistance gene blaNDM (p < 0.0001). Virulence genes such as mrkD (97.6%), uge (96.9%), kpn (96.9%), and fim-H (84.3%) had high detection rates. IncF (57.5%) was the major replicon plasmid detected, and sequencing showed that the CRKP063 genome contained two plasmids. The plasmid carrying blaKPC-2, which mediates carbapenem resistance, was located on the 359,625 base pair plasmid IncFII, together with virulence factors, plasmid replication protein (rep B), stabilizing protein (par A), and type IV secretion system (T4SS) proteins that mediate plasmid conjugation transfer.ConclusionOur study aids in understanding the prevalence of CRKP in this hospital and the significant differences between children and adults, thus providing new ideas for clinical empirical use of antibiotics.

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