Use of repetitive sequence-based polymerase chain reaction for molecular epidemiologic analysis of Streptococcus equi subspecies equi.
Abstract: To determine whether repetitive sequence-based polymerase chain reaction (rep-PCR) could be used to differentiate Streptococcus equi isolates, to examine S equi isolates from throughout the world, and to determine whether a horse had > 1 subtype of S equi during an outbreak of disease. Methods: An initial group of 32 S equi isolates, 63 S equi isolates from various geographic areas, and 17 S equi isolates obtained during outbreaks of disease. Methods: An aliquot of S equi genomic DNA was amplified, using enterobacterial repetitive intergenic consensus primers. Gel electrophoresis was performed on 1.5% agarose gels, and a computed-assisted program was used to compare rep-PCR results. Results: Use of these primers to analyze 100 ng of S equi genomic DNA resulted in patterns of 6 to 14 bands. The 32 initial isolates were separated into 7 rep-PCR subtypes. There were 30 rep-PCR subtypes found among 29 S equi isolates obtained from Minnesota, Michigan, Canada, and Australia and 34 S equi isolates obtained from Kentucky and other sources. Furthermore, the same clone was identified in several horses during an outbreak of disease. Infected horses on the same farm all had a single clone of S equi. Conclusions: Analysis of these results suggests that rep-PCR is useful for delineating S equi into rep-PCR subtypes. Results revealed that isolates with the same geographic source or similar date of collection did not always have the same rep-PCR subtype. A single clone of S equi usually predominated during an outbreak of disease.
Publication Date: 2000-06-13 PubMed ID: 10850848DOI: 10.2460/ajvr.2000.61.699Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article is an investigation on the use of repetitive sequence-based polymerase chain reaction (rep-PCR) in distinguishing different Streptococcus equi isolates, their geographical distribution, and prevalence in horse disease outbreaks.
Objective and Methods of the Study
- The study aimed to explore the utility of rep-PCR in differentiating Streptococcus equi isolates, to analyze a variety of S equi isolates from across the globe, and to ascertain whether a horse could carry more than one S equi subtype during an outbreak.
- The researchers initially worked with 32 S equi isolates, extending their study to 63 isolates with various geographies, and later focusing on 17 isolates discovered during disease outbreaks.
- The method employed involved the amplification of an aliquot of S equi genomic DNA using enterobacterial repetitive intergenic consensus primers.
- Gel electrophoresis was performed on the amplified DNA and a computer-assisted program was used to compare the resulting rep-PCR patterns.
Results and Findings
- The use of the specific primers to analyze S equi revealed varied patterns of bands ranging from 6 to 14, upon gel electrophoresis. This suggested different S equi subtypes.
- The initial 32 isolates branched into 7 rep-PCR subtypes.
- Among 29 S equi isolates sourced from Minnesota, Michigan, Canada, and Australia, and 34 isolates taken from Kentucky and other places, 30 rep-PCR subtypes were found.
- During an outbreak, several horses were found to carry the same clone of S equi, indicating that a single S equi clone usually dominated an outbreak.
Conclusion of the Study
- The research concluded that rep-PCR is a useful tool for categorizing S equi into various rep-PCR subtypes.
- The outcomes revealed that S equi isolates carrying the same geographic source or a close collection date did not necessarily belong to the same rep-PCR subtype.
- The study further confirmed that typically, a single clone of S equi prevailed during a disease outbreak.
Cite This Article
APA
Al-Ghamdi GM, Kapur V, Ames TR, Timoney JF, Love DN, Mellencamp MA.
(2000).
Use of repetitive sequence-based polymerase chain reaction for molecular epidemiologic analysis of Streptococcus equi subspecies equi.
Am J Vet Res, 61(6), 699-705.
https://doi.org/10.2460/ajvr.2000.61.699 Publication
Researcher Affiliations
- Department of Clinical and Population Sciences, College of Veterinary Medicine, University of Minnesota, St Paul 55108, USA.
MeSH Terms
- Animals
- Australia / epidemiology
- DNA Fingerprinting / veterinary
- DNA Primers / chemistry
- DNA, Bacterial / chemistry
- DNA, Bacterial / isolation & purification
- Disease Outbreaks / classification
- Disease Outbreaks / veterinary
- Electrophoresis, Agar Gel / veterinary
- Europe / epidemiology
- Horse Diseases / epidemiology
- Horse Diseases / microbiology
- Horses
- Midwestern United States / epidemiology
- Ontario / epidemiology
- Phylogeny
- Polymerase Chain Reaction / veterinary
- Streptococcal Infections / epidemiology
- Streptococcal Infections / microbiology
- Streptococcal Infections / veterinary
- Streptococcus equi / chemistry
- Streptococcus equi / classification
- Streptococcus equi / genetics
Citations
This article has been cited 2 times.- Sá e Silva M, da Costa MM, de Avila Botton S, Barretta C, Groff AC, de Vargas AC. Phenotypical assays and partial sequencing of the hsp60 gene for identification of Streptococcus equi. Curr Microbiol 2007 May;54(5):331-4.
- Kelly C, Bugg M, Robinson C, Mitchell Z, Davis-Poynter N, Newton JR, Jolley KA, Maiden MC, Waller AS. Sequence variation of the SeM gene of Streptococcus equi allows discrimination of the source of strangles outbreaks. J Clin Microbiol 2006 Feb;44(2):480-6.
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