Evaluation of (GTG) 5-PCR for Genotyping of Klebsiella pneumonia Strains Isolated from Patients with Urinary Tract Infections

Ranjbar, Reza; Afshar, Davoud

doi:10.18502/ijph.v48i10.3496

Cited by 7 publications

(7 citation statements)

References 22 publications

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“…Klebsiella pneumoniae is a gram negative and rodshaped bacillus from the genus Klebsiella and family Enterobacteriaceae. As a nosocomial pathogen, it causes several serious infections in-cluding pneumonia, bacteremia, hepatic abscesses and urinary tract infections particularly in the patients with underlying disease such as diabetes mellitus (1,2). Because of rapid emergence and spread of antibiotic resistance between the bacteria, there is an urgent need to monitor the use of antimicrobial agents and modify antibiotic therapies.…”

Section: Introductionmentioning

confidence: 99%

“…Klebsiella pneumoniae is a gram negative and rod-shaped bacillus from the genus Klebsiella and family Enterobacteriaceae. As a nosocomial pathogen, it causes several serious infections including pneumonia, bacteremia, hepatic abscesses and urinary tract infections particularly in the patients with underlying disease such as diabetes mellitus ( 1 , 2 ).…”

Section: Introductionmentioning

confidence: 99%

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Molecular Detection of gyrA Mutation in Clinical Strains of Klebsiella pneumoniae

Ahmadi¹,

Noormohammadi²,

Behzadi³

et al. 2022

ijph

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Background: Multi-drug resistant (MDR) Klebsiella pneumoniae strains cause the majority of community acquired and life-threatening infections. We aimed to detect the gyrA mutations in the clinical strains of nalidixic acid and ciprofloxacin resistant K. pneumoniae isolated from the patients with urinary tract infections. Methods: Bacterial strains were isolated from the patients with urinary tract infections admitted to a major hospital in Tehran, Iran (2017-2018). Bacterial identification was done according to standard microbiological tests. Antimicrobial susceptibility testing for quinolones and fluoroquinolone antibiotics was done using both disc diffusion and minimal inhibitory concentrations (MICs) methods. PCR-RFLP was used to detect the probable mutation in the gyrA gene in nalidixic acid and ciprofloxacin resistant strains. Finally sequencing was performed to detect point mutations in isolated K. pneumoniae strains. Results: One hundred K. pneumonia isolates were recovered from the urine samples of the clinical cases. Antibiotic resistance testing showed that among all K. pneumoniae isolates, 26% and 19% of the strains were resistant to nalidixic acid and ciprofloxacin respectively. MIC value was ≥4 µg/ml for ciprofloxacin resistant isolates. The results of RFLP on gyrA PCR amplicons using HinfI restriction enzyme showed point mutation in this gene in 46% of nalidixic acid and ciprofloxacin resistant K. pneumonia. The data obtained from the sequencing confirmed the RFLP results and indicated the presence of point mutations in codons 83 and 87 in the gyrA gene which leads to the substitution of different amino acids in gyrA protein. Conclusion: Our findings indicated a relative increased rate of resistance against quinolones and fluoroquinolone antibiotics that raised a concern about extensive dissemination of clinical strains of nalidixic acid and ciprofloxacin resistant K. pneumonia. Point mutation of gyrA gene was responsible for the resistance in our strains however to gain more insight into the molecular characterization of quinolone-resistant isolates, other possible mechanisms of the resistance should also be investigated.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Molecular Detection of gyrA Mutation in Clinical Strains of Klebsiella pneumoniae

Ahmadi¹,

Noormohammadi²,

Behzadi³

et al. 2022

ijph

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“…The presence of DNA was assessed by qualitative analysis in agarose gel electrophoresis. The supernatant was stored at -20° C for performing PCR [18].…”

Section: Molecular Characterization By (Gtg) 5 -Pcr Dna Extractionmentioning

confidence: 99%

“…khare et al and Mohapatra et al had used the same method to grouping E. coli strains [8,32], similarly Ranjbar and. Afshar had grouped together the strains of uropathogenic Klebsiella pneumoniae by the same method [18]. According to molecular analysis and phylogeny tree based profile of fingerprinting of strains, we find that the strains of groups G13 (P66), G10 (P80) and G19 (P46) isolated from pus could also be the cause of urinary tract infections due to of their affiliation at strains responsible of urinary pathologies according to Fig.…”

Section: Phylogenetic Affiliationmentioning

confidence: 99%

(GTG)5-PCR fingerprinting of multi-drug resistant Escherichia coli bacteria isolates from hospital in Ouagadougou, Burkina Faso

et al. 2022

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Background Escherichia coli (E. coli) is the most common bacterial species implicated in various types of infections including septicemia, gastroenteritis, urinary tract infections, meningitis and others pathologies. These involve several bacterial clones with multidrug resistance making them difficult to treat. The aims of this study was to perform molecular typing of E. coli strains using universal primer (GTG)5. In this study, 53 E. coli strains were collected from inpatients and outpatients. The test of antimicrobial sensibility was realized using CA-SFM /EUCAST method and strains were identified by conventional microbiological tests. The carbapenemase-producing strains were demonstrated by phenotypic method. Bacterial DNA was extracted by boiling method. (GTG)5-PCR was used for strain subtyping. The DendroUPGMA software was used for grouping of strains from the genetic fingerprints obtained by (GTG)5-PCR. Results Antibiotic susceptibility test revealed that all strains were multi-drug resistant (MDR). Its strains showed resistance to at least three different families of antibiotics. Of this MDR strains, only one was a metallo-β-lactamase producer. The dendrogram obtained using genetic fingerprinting allowed the E. coli strains to be grouped into 22 clusters (G1 to G22). Conclusion The (GTG) 5-PCR assay enabled rapid molecular typing of E. coli strains. The strains of E. coli typed in this study would belong to different clones.

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“…The pathogenicity of this organism is notable because of its ability to produce some important enzymes which destroy an extended spectrum of antimicrobial agents (Shakib et al, 2012). In a study, 88 K. pneumoniae isolates were subjected to (GTG) 5 -PCR assay, and the strains were differentiated into 9 clusters which showed the ability of this method for molecular typing of K. pneumoniae strains (Ranjbar and Afshar, 2019). Svec et al (2008) evaluated the (GTG) 5 -PCR for the fast screening of bacterial strains isolated from dental plaque of children with early childhood caries.…”

Section: Klebsiella Speciesmentioning

confidence: 99%

(GTG)5-PCR Mediated Molecular Typing of Zoonotic Bacteria

Babazadeh¹,

Ranjbar²

2021

WVJ

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The present review aimed to reveal the role of (GTG)5-PCR microbial typing in indicating the routes and source of infections, investigate the outbreaks and genotypes of clinical strains, as well as finding virulent strains and epidemiology of bacterial isolates. All available and published data in Google scholar, PubMed, ResearchGate, and Science Direct during the past two decades that used the (GTG)5-PCR method for genotyping the bacterial isolates were included in the current study. The findings have indicated that (GTG)5-PCR can be recommended as a possible, cost-effective, fast, and easy tool for molecular typing of bacterial isolates.

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Evaluation of (GTG) 5-PCR for Genotyping of Klebsiella pneumonia Strains Isolated from Patients with Urinary Tract Infections

Cited by 7 publications

References 22 publications

Molecular Detection of gyrA Mutation in Clinical Strains of Klebsiella pneumoniae

Molecular Detection of gyrA Mutation in Clinical Strains of Klebsiella pneumoniae

(GTG)5-PCR fingerprinting of multi-drug resistant Escherichia coli bacteria isolates from hospital in Ouagadougou, Burkina Faso

(GTG)5-PCR Mediated Molecular Typing of Zoonotic Bacteria

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