Comparison of (GTG)5-oligonucleotide and ribosomal intergenic transcribed spacer (ITS)-PCR for molecular typing of Klebsiella isolates

Ryberg, Anna; Olsson, Cecilia; Ahrné, Siv; Monstein, Hans‐Jürg

doi:10.1016/j.mimet.2010.11.019

Cited by 13 publications

(9 citation statements)

References 34 publications

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“…(GTG) 5 oligonucleotide typing was performed as previously described (Ryberg et al, 2011 ) with minor modifications. The amplification reaction was carried out in a total volume of 25 μL consisting of 1.0 U of Taq DNA polymerase (Dongsheng Biotech,Guangzhou, China), 1.0 μM of primer, 2.5 mM MgCl 2 , 0.25 mM of each dNTP, and 40 ng of template genomic DNA.…”

Section: Methodsmentioning

confidence: 99%

“…This is particularly important in endemic and epidemic nosocomial outbreaks of K. pneumoniae infections to improve the management of such outbreaks. A variety of methods have been used for K. pneumoniae typing, including pulsed-field gel electrophoresis (PFGE), enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR), randomly amplified polymorphic DNA (RAPD), and (GTG) 5 oligonucleotide PCR (Haryani et al, 2007 ; Ryberg et al, 2011 ; Barus et al, 2013 ; Sachse et al, 2014 ). The ERIC, RAPD, and (GTG) 5 -PCR assays are relatively simple and cost-effective methods that have been successfully used for genotyping K. pneumoniae isolates from various sources (Ryberg et al, 2011 ; Barus et al, 2013 ).…”

Section: Introductionmentioning

confidence: 99%

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Phenotypic and Genotypic Characterization of Klebsiella pneumoniae Isolated From Retail Foods in China

Zhang

Yang

et al. 2018

Front. Microbiol.

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Klebsiella pneumoniae is not only a major hospital-acquired pathogen but also an important food-borne pathogen that can cause septicaemia, liver abscesses, and diarrhea in humans. The phenotypic and genotypic characteristics of K. pneumoniae in retail foods have not been thoroughly investigated in China. The objective of this study was to characterize K. pneumoniae isolates through biotyping, serotyping, determination of virulence factors, antibiotic resistance testing, enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR), and (GTG)5-PCR molecular typing. From May 2013 to April 2014, a total of 61 K. pneumoniae isolates were collected from retail foods in China. Using API 20E test strips, five different biotype profiles were identified among these isolates. The majority of isolates belonged to biochemical profile “5215773” (50 isolates, 80.6%). The capsular serotypes of the 61 K. pneumoniae isolates and one reference strain were determined by PCR. Of the seven capsular serotypes tested, four different capsular serotypes were identified. Serotypes K1, K20, K57, and K2 were detected in two, three, two, and one isolates, respectively. Serotypes K3, K5, and K54 were not detected. The presence of 11 virulence genes was assessed by PCR. The most common virulence genes were fimH (85.5%), ureA (79.0%), wabG (77.4%), uge (56.5%), and kfuBC (29.0%). ERIC-PCR and (GTG)5-PCR molecular typing indicated high genetic diversity among K. pneumoniae isolates. We identified 60 different ERIC patterns and 56 distinct (GTG)5 patterns. Genotypic results indicated that isolates carrying similar virulence factors were generally genetically related. Some isolates from the same geographic area have a closer relationship. The isolates showed high levels of resistance to ampicillin (51/62, 82.2%). Resistance to streptomycin (11/62, 17.7%) and piperacillin (10/62, 16.1%) was also common. The presence of virulent and antibiotic-resistant K. pneumoniae in foods poses a potential health hazard for consumers. Our findings highlight the importance of surveillance of K. pneumoniae in foods.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Phenotypic and Genotypic Characterization of Klebsiella pneumoniae Isolated From Retail Foods in China

Zhang

Yang

et al. 2018

Front. Microbiol.

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“…The GTG-PCR was chosen as it was successfully employed in our laboratory for inter-serovar discrimination of Salmonella strains [ 15 ]. What is more, (GTG) 5 -PCR tests were used for genotyping Klebsiella strains [ 20 ] and identification of Streptococcus mutans , Staphylococcus spp. and Enterococcus spp [ 21 – 23 ].…”

Section: Discussionmentioning

confidence: 99%

TRS-PCR profiling for discrimination of Escherichia coli strains isolated from children with diarrhea under 5 years of age in Lodz region, Poland

Kubiak-Szeligowska

Bartnicka

Jarych³

et al. 2016

Mol Biol Rep

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Escherichia coli is one of the most frequently isolated gram-negative pathogens in cases of foodborne diseases and hospital infections. What is more, diarrheal diseases, including these associated with pathogenic E. coli strains, are leading causes of morbidity and mortality worldwide, especially among children. Improvements of the management of diarrheal diseases caused by these bacteria are in the spotlight of the World Health Organization. Therefore, there is still a need to develop new methods or improve ones that are commonly used to characterize and distinguish E. coli strains more precisely. In this work, TRS-based PCRs were effectively used for discrimination of 123 E. coli strains isolated from children with diarrhea in the Lodz region (Poland). The composite TRS-PCR approach, based on similarity comparisons of GTG-PCR and CGG-PCR fingerprints, enabled us to distinguish strains with very good efficacy. This was confirmed by the high diversity index (0.991) and high reproducibility of the band patterns obtained (95.0 %). These results showed the great variation in strains that may cause infections in children under 38 months. However, the stains were grouped in three separate clusters, which were different in terms of their phylogenetic affiliation and virulence factor repertoire. The obtained results support and are consistent with the need of public health surveillance for searching new and fast assays as far as children’s health is concerned. TRS-PCR profiling is an effective tool for genotyping of E. coli strains isolated from children with diarrhea.

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“…Thus, Ryberg et al [60] applied poly-trinucleotide (GTG) 5 and rDNA ITS-PCR-CE fingerprint analysis to genotype Klebsiella oxytoca and Klebsiella pneumoniae isolates. Dendrograms were constructed and the results indicated that K. oxytoca and K. pneumoniae isolates, reference strains, and standard strains formed disparate cluster groups, and tentative subclusters were built, indicating that (GTG) 5 and ITS-PCR-CE analysis is a promising tool for genotyping Klebsiella isolates.…”

Section: Detecting and Diagnosing Bacteriamentioning

confidence: 99%

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Lian

Zhao

2016

Clinical Chemistry and Laboratory Medicine (CCLM)

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Rapid transmission, high morbidity, and mortality are the features of human infectious diseases caused by microorganisms, such as bacteria, fungi, and viruses. These diseases may lead within a short period of time to great personal and property losses, especially in regions where sanitation is poor. Thus, rapid diagnoses are vital for the prevention and therapeutic intervention of human infectious diseases. Several conventional methods are often used to diagnose infectious diseases, e.g. methods based on cultures or morphology, or biochemical tests based on metabonomics. Although traditional methods are considered gold standards and are used most frequently, they are laborious, time consuming, and tedious and cannot meet the demand for rapid diagnoses. Disease diagnosis using capillary electrophoresis methods has the advantages of high efficiency, high throughput, and high speed, and coupled with the different nucleic acid detection strategies overcomes the drawbacks of traditional identification methods, precluding many types of false positive and negative results. Therefore, this review focuses on the application of capillary electrophoresis based on nucleic detection to the diagnosis of human infectious diseases, and offers an introduction to the limitations, advantages, and future developments of this approach.

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Comparison of (GTG)5-oligonucleotide and ribosomal intergenic transcribed spacer (ITS)-PCR for molecular typing of Klebsiella isolates

Abstract: Molecular typing of Klebsiella species has become important for monitoring dissemination of β-lactamase-producers in hospital environments. The present study was designed to evaluate poly-trinucleotide (GTG) 5

Cited by 13 publications

References 34 publications

Phenotypic and Genotypic Characterization of Klebsiella pneumoniae Isolated From Retail Foods in China

Phenotypic and Genotypic Characterization of Klebsiella pneumoniae Isolated From Retail Foods in China

TRS-PCR profiling for discrimination of Escherichia coli strains isolated from children with diarrhea under 5 years of age in Lodz region, Poland

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

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