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 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 Applicability of Three Alternative Instruments for Food Authenticity Analysis:GMO Identification

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
A. Burrell ◽  
C. Foy ◽  
M. Burns
Keyword(s):  
Polymerase Chain Reaction ◽  
Gel Electrophoresis ◽  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
Fitness For Purpose ◽  
Chain Reaction ◽  
Food Authenticity ◽  
Pcr Products ◽  
Polymerase Chain

Ensuring foods are correctly labelled for ingredients derived from genetically modified organisms (GMOs) is an issue facing manufacturers, retailers, and enforcement agencies. DNA approaches for the determination of food authenticitys often use the polymerase chain reaction (PCR), and PCR products can be detected using capillary or gel electrophoresis. This study examines the fitness for purpose of the application of three laboratory electrophoresis instruments (Agilent Bioanalyzer 2100, Lab901 TapeStation, and Shimadzu MCE-202 MultiNA) for the detection of GMOs using PCR based on a previously validated protocol. Whilst minor differences in the performance characteristics of bias and precision were observed, all three instruments demonstrated their applicability in using this protocol for screening of GMO ingredients.

Gel electrophoresis of polymerase chain reaction (PCR) products improves GenoType® MRSA Direct PCR analysis in clinical practice

2008 ◽  
Vol 68 (3) ◽  
pp. 276-277
Author(s):  
S. Borgmann ◽  
H. Gruber ◽  
K. Beyser ◽  
R. Abel ◽  
A. Reber ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Clinical Practice ◽  
Gel Electrophoresis ◽  
Pcr Analysis ◽  
Chain Reaction ◽  
Direct Pcr ◽  
Pcr Products ◽  
Polymerase Chain

scholarly journals Expression of 5’-deiodinase in various thyroid tumors

2004 ◽  
Vol 50 (3) ◽  
pp. 34-37
Author(s):  
G. M. Artykbaeva ◽  
Ya. Kh Turakulov
Keyword(s):  
Polymerase Chain Reaction ◽  
Thyroid Carcinoma ◽  
Gel Electrophoresis ◽  
Human Thyroid ◽  
Thyroid Tumors ◽  
Great Role ◽  
Chain Reaction ◽  
Pcr Products ◽  
Important Pathway ◽  
Polymerase Chain

5'-deiodination is an important pathway for the metabolism of T4 by determining its great role as a prohormone for T3. T4 is metabolized by two 5'-deiodinases: DI and DII which are encoded by different genes, regulated by different modes and expressed in various tissues. The aim of the present study was to examine DI and DII expression in health and in various human thyroid tumors. Eighteen thyroid specimens (10 from tumors and 8 from the intact thyroid tissues surrounding the nodes) were used. RNA isolated from the tissues by reserve transcription, followed by polymerase chain reaction (PCR) were transcribed to cDNA. Gel electrophoresis showed the bands corresponding to 177 b. p for DI and 796 b. p for DII. The experiments demonstrated that there were genes of both isoenzymes of 5 ’deiodinases in thyroid carcinoma tissues. Gel electrophoresis indicated the different expression of PCR products in different thyroid tumors for DI and DII at the level of mRNA.

scholarly journals A Comparative Study of 5S rDNA Non-Transcribed Spacers in Elaeagnaceae Species

Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 4
Author(s):  
Oleg S. Alexandrov ◽  
Olga V. Razumova ◽  
Gennady I. Karlov
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Markers ◽  
Dna Markers ◽  
5S Rdna ◽  
Chain Reaction ◽  
Closely Related Species ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Species Specific ◽  
Shepherdia Canadensis

5S rDNA is organized as a cluster of tandemly repeated monomers that consist of the conservative 120 bp coding part and non-transcribed spacers (NTSs) with different lengths and sequences among different species. The polymorphism in the 5S rDNA NTSs of closely related species is interesting for phylogenetic and evolutional investigations, as well as for the development of molecular markers. In this study, the 5S rDNA NTSs were amplified with universal 5S1/5S2 primers in some species of the Elaeagnaceae Adans. family. The polymerase chain reaction (PCR) products of five Elaeagnus species had similar lengths near 310 bp and were different from Shepherdia canadensis (L.) Nutt. and Sh. argentea (Pusch.) Nutt. samples (260 bp and 215 bp, respectively). The PCR products were cloned and sequenced. An analysis of the sequences revealed that intraspecific levels of NTS identity are high (approximately 95–96%) and similar in the Elaeagnus L. species. In Sh. argentea, this level was slightly lower due to the differences in the poly-T region. Moreover, the intergeneric and intervarietal NTS identity levels were studied and compared. Significant differences between species (except E. multiflora Thunb. and E. umbellata Thunb.) and genera were found. Herein, a range of the NTS features is discussed. This study is another step in the investigation of the molecular evolution of Elaeagnaceae and may be useful for the development of species-specific DNA markers in this family.

scholarly journals 484. DNA analysis with polymerase chain reaction (PCR) of bacterial infections

1992 ◽  
Vol 48 (8) ◽  
pp. 1567
Author(s):  
Takashi Arai ◽  
Hideaki Yamamoto ◽  
Satoshi Kobayashi ◽  
Akira Yamaguchi ◽  
Katsumi Ikei
Keyword(s):  
Polymerase Chain Reaction ◽  
Bacterial Infections ◽  
Dna Analysis ◽  
Chain Reaction ◽  
Polymerase Chain
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