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Veterinary research communications2001; 25(4); 261-269; doi: 10.1023/a:1010674524428

Comparison of the value of pulsed-field gel electrophoresis, random amplified polymorphic DNA and amplified rDNA restriction analysis for subtyping Taylorella equigenitalis.

Abstract: Eight strains of Taylorella equigenitalis were identified by a polymerase chain reaction using a primer pair specific to the 16S rDNA of T equigenitalis. These eight strains were chosen because they had previously been shown to represent eight distinct genotypes by pulsed-field gel electrophoresis analysis after separate digestion of the genomic DNA with ApaI or NotI. The eight strains could be classified into six or seven types by random amplified polymorphic DNA analysis using different kinds of primers. Amplified rDNA restriction analysis after separate digestion with five restriction enzymes, including AluI and MboI, of the 1,500 bp fragments of rDNA amplified by polymerase chain reaction did not discriminate the genomic variations among the eight strains of T equigenitalis. Thus, pulsed-field gel electrophoresis was shown to discriminate these eight organisms better than random amplified polymorphic DNA analysis, while amplified rDNA restriction analysis was found to be unsuitable for subtyping T equigenitalis.
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Publication Date: 2001-07-04 PubMed ID: 11432428DOI: 10.1023/a:1010674524428Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research compares the effectiveness of three genetic analysis methods – pulsed-field gel electrophoresis, random amplified polymorphic DNA analysis, and amplified rDNA restriction analysis – in differentiating eight strains of Taylorella equigenitalis, a bacterium. The study found that pulsed-field gel electrophoresis was the most effective procedure, while amplified rDNA restriction analysis was deemed unsuitable.

Research Methodology

  • The study focused on eight strains of Taylorella equigenitalis. These strains were selected because previous research had confirmed their differentiation into eight unique genotypes through pulsed-field gel electrophoresis (PFGE) following separate digestion of the DNA with two enzymes (ApaI or NotI).
  • The scientists then employed a polymerase chain reaction (PCR) using a primer pair specific to the 16S rDNA of T equigenitalis to identify the strains.

Comparison of Analysis Techniques

  • The first technique tested was random amplified polymorphic DNA (RAPD) analysis, which classified the eight strains into six or seven types, depending on the primers used.
  • The second technique, amplified rDNA restriction analysis, entailed the separation digestion with five restriction enzymes, including AluI and MboI. This technique was performed on 1,500 base pairs fragments of rDNA that were amplified by PCR. This method, however, did not reveal the genomic variations among the eight strains of T. equigenitalis.
  • The third method, previously mentioned PFGE, demonstrated the ability to most effectively differentiate the eight T. equigenitalis strains.

Findings and Conclusion

  • The study concluded that pulsed-field gel electrophoresis was the most effective procedure for subtyping the eight strains of Taylorella equigenitalis, outperforming random amplified polymorphic DNA analysis.
  • On the other hand, amplified rDNA restriction analysis was found to be unsuitable for this purpose because it failed to distinguish the genomic variations in the strains. This highlighted the necessity of choosing the appropriate analysis method for accurate differentiation and subtyping of bacterial strains.

Cite This Article

APA
Kagawa S, Moore JE, Murayama O, Matsuda M. (2001). Comparison of the value of pulsed-field gel electrophoresis, random amplified polymorphic DNA and amplified rDNA restriction analysis for subtyping Taylorella equigenitalis. Vet Res Commun, 25(4), 261-269. https://doi.org/10.1023/a:1010674524428

Publication

Veterinary research communications
ISSN: 0165-7380
NlmUniqueID: 8100520
Country: Switzerland
Language: English
Volume: 25
Issue: 4
Pages: 261-269

Researcher Affiliations

Kagawa, S
  • Laboratory of Molecular Biology, Graduate School of Environmental Health Sciences, Azabu University, Sagamihara, Japan.
Moore, J E
    Murayama, O
      Matsuda, M

        MeSH Terms

        • Animals
        • DNA, Bacterial / chemistry
        • DNA, Bacterial / genetics
        • DNA, Bacterial / isolation & purification
        • DNA, Ribosomal / chemistry
        • DNA, Ribosomal / genetics
        • DNA, Ribosomal / isolation & purification
        • Electrophoresis, Gel, Pulsed-Field / methods
        • Electrophoresis, Gel, Pulsed-Field / veterinary
        • Gram-Negative Bacterial Infections / diagnosis
        • Gram-Negative Bacterial Infections / microbiology
        • Gram-Negative Bacterial Infections / veterinary
        • Horse Diseases / diagnosis
        • Horse Diseases / microbiology
        • Horses
        • Polymerase Chain Reaction / veterinary
        • Random Amplified Polymorphic DNA Technique / methods
        • Random Amplified Polymorphic DNA Technique / veterinary
        • Restriction Mapping / veterinary
        • Taylorella equigenitalis / chemistry
        • Taylorella equigenitalis / classification
        • Taylorella equigenitalis / genetics

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        Citations

        This article has been cited 2 times.
        1. Hicks J, Stuber T, Lantz K, Erdman M, Robbe-Austerman S, Huang X. Genomic diversity of Taylorella equigenitalis introduced into the United States from 1978 to 2012. PLoS One 2018;13(3):e0194253.
          doi: 10.1371/journal.pone.0194253pubmed: 29584782google scholar: lookup
        2. Matsuda M, Tazumi A, Kagawa S, Sekizuka T, Murayama O, Moore JE, Millar BC. Homogeneity of the 16S rDNA sequence among geographically disparate isolates of Taylorella equigenitalis. BMC Vet Res 2006 Jan 6;2:1.
          doi: 10.1186/1746-6148-2-1pubmed: 16398935google scholar: lookup
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