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.
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
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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
Researcher Affiliations
- Laboratory of Molecular Biology, Graduate School of Environmental Health Sciences, Azabu University, Sagamihara, Japan.
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
References
This article includes 23 references
- Bleumink-Pluym N, ter Laak EA, van der Zeijst BA. Epidemiologic study of Taylorella equigenitalis strains by field inversion gel electrophoresis of genomic restriction endonuclease fragments.. J Clin Microbiol 1990 Sep;28(9):2012-6.
- Matsuda M, Miyazawa T, Moore JE, Buckley TC, Thomas LA. Molecular genotyping by pulsed-field gel electrophoresis of restricted genomic DNA of strains of Taylorella equigenitalis isolated in Ireland and in the United States.. Vet Res Commun 1998 Jun;22(4):217-24.
- Crowhurst RC. Genital infection in mares.. Vet Rec 1977 May 28;100(22):476.
- van Belkum A, Bax R, Peerbooms P, Goessens WH, van Leeuwen N, Quint WG. Comparison of phage typing and DNA fingerprinting by polymerase chain reaction for discrimination of methicillin-resistant Staphylococcus aureus strains.. J Clin Microbiol 1993 Apr;31(4):798-803.
- Endtz HP, Vliegenthart JS, Vandamme P, Weverink HW, van den Braak NP, Verbrugh HA, van Belkum A. Genotypic diversity of Campylobacter lari isolated from mussels and oysters in The Netherlands.. Int J Food Microbiol 1997 Jan;34(1):79-88.
- Akopyanz N, Bukanov NO, Westblom TU, Kresovich S, Berg DE. DNA diversity among clinical isolates of Helicobacter pylori detected by PCR-based RAPD fingerprinting.. Nucleic Acids Res 1992 Oct 11;20(19):5137-42.
- Liebisch B, Schwarz S. Evaluation and comparison of molecular techniques for epidemiological typing of Salmonella enterica subsp. enterica serovar dublin.. J Clin Microbiol 1996 Mar;34(3):641-6.
- Old DC, Rankin SC, Crichton PB. Assessment of strain relatedness among Salmonella serotypes Salinatis, Duisburg, and Sandiego by biotyping, ribotyping, IS200 fingerprinting, and pulsed-field gel electrophoresis.. J Clin Microbiol 1999 Jun;37(6):1687-92.
- Matsuda M, Kagawa S, Sakamoto Y, Miyajima M, Barton M, Moore JE. Detection of heterogeneous genotypes among Australian strains of Taylorella equigenitalis.. Aust Vet J 2000 Jan;78(1):56-7.
- Weisburg WG, Barns SM, Pelletier DA, Lane DJ. 16S ribosomal DNA amplification for phylogenetic study.. J Bacteriol 1991 Jan;173(2):697-703.
- Taylor CE, Rosenthal RO, Brown DF, Lapage SP, Hill LR, Legros RM. The causative organism of contagious equine metritis 1977: proposal for a new species to be known as Haemophilus equigenitalis.. Equine Vet J 1978 Jul;10(3):136-44.
- Savor C, Pfaller MA, Kruszynski JA, Hollis RJ, Noskin GA, Peterson LR. Comparison of genomic methods for differentiating strains of Enterococcus faecium: assessment using clinical epidemiologic data.. J Clin Microbiol 1998 Nov;36(11):3327-31.
- van der Zee A, Verbakel H, van Zon JC, Frenay I, van Belkum A, Peeters M, Buiting A, Bergmans A. Molecular genotyping of Staphylococcus aureus strains: comparison of repetitive element sequence-based PCR with various typing methods and isolation of a novel epidemicity marker.. J Clin Microbiol 1999 Feb;37(2):342-9.
- Chetoui H, Melin P, Struelens MJ, Delhalle E, Nigo MM, De Ryck R, De Mol P. Comparison of biotyping, ribotyping, and pulsed-field gel electrophoresis for investigation of a common-source outbreak of Burkholderia pickettii bacteremia.. J Clin Microbiol 1997 Jun;35(6):1398-403.
- Wada S, Matsuda M, Kikuchi M, Kodama T, Takei I, Ogawa S, Takahashi S, Shingaki M, Itoh T. Genome DNA analysis and genotyping of clinical isolates of Helicobacter pylori.. Cytobios 1994;80(321):109-16.
- ter Laak EA, Fennema G, Jaartsveld FH. [Contagious equine metritis in The Netherlands].. Tijdschr Diergeneeskd 1989 Feb 15;114(4):189-201.
- Miyazawa T, Matsuda M, Isayama Y, Samata T, Ishida Y, Ogawa S, Takei K, Honda M, Kamada M. Genotyping of isolates of Taylorella equigenitalis from thoroughbred brood mares in Japan.. Vet Res Commun 1995;19(4):265-71.
- Timoney PJ, Strickland KL. CEM (contagious equine metritis) in the Republic of Ireland.. Vet Rec 1982 Oct 23;111(17):400-1.
- Miserez R, Frey J, Krawinkler M, Nicolet J. [Identification and diagnosis of Taylorella equigenitalis by a DNA amplification method (PCR)].. Schweiz Arch Tierheilkd 1996;138(3):115-20.
- Matsuda M, Miyazawa T, Ishida Y, Moore JE. Profiles of fragments after pulsed-field gel electrophoresis of cleaved genomic DNA from strains of Taylorella equigenitalis isolated from horses in Norway.. Microbiol Res 1997 Jul;152(2):217-20.
- Gomez-Lus P, Fields BS, Benson RF, Martin WT, O'Connor SP, Black CM. Comparison of arbitrarily primed polymerase chain reaction, ribotyping, and monoclonal antibody analysis for subtyping Legionella pneumophila serogroup 1.. J Clin Microbiol 1993 Jul;31(7):1940-2.
- Timoney PJ, Powell DG. Isolation of the contagious equine metritis organism from colts and fillies in the United Kingdom and Ireland.. Vet Rec 1982 Nov 20;111(21):478-82.
- Rafferty ME, Baltch AL, Smith RP, Bopp LH, Rheal C, Tenover FC, Killgore GE, Lyerly DM, Wilkins TD, Schoonmaker DJ, Hannett GE, Shayegani M. Comparison of restriction enzyme analysis, arbitrarily primed PCR, and protein profile analysis typing for epidemiologic investigation of an ongoing Clostridium difficile outbreak.. J Clin Microbiol 1998 Oct;36(10):2957-63.
Citations
This article has been cited 2 times.- 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.
- 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.
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