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Home / Equine Research Bank / Vet Microbiol / Real-time PCR for detection and differentiation of Streptoco...
Veterinary microbiology2007; 124(3-4); 219-229; doi: 10.1016/j.vetmic.2007.04.020

Real-time PCR for detection and differentiation of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus.

Abstract: Strangles is a contagious equine disease caused by Streptococcus equi subsp. equi. In this study, clinical strains of S. equi (n=24) and Streptococcus equi subsp. zooepidemicus (n=24) were genetically characterized by sequencing of the 16S rRNA and sodA genes in order to devise a real-time PCR system that can detect S. equi and S. zooepidemicus and distinguish between them. Sequencing demonstrated that all S. equi strains had the same 16S rRNA sequence, whereas S. zooepidemicus strains could be divided into subgroups. One of these (n=12 strains) had 16S rRNA sequences almost identical with the S. equi strains. Interestingly, four of the strains biochemically identified as S. zooepidemicus were found by sequencing of the 16S rRNA gene to have a sequence homologous with Streptococcus equi subsp. ruminatorum. However, they did not have the colony appearance or the biochemical characteristics of the type strain of S. ruminatorum. Classification of S. ruminatorum may thus not be determined solely by 16S rRNA sequencing. Sequencing of the sodA gene demonstrated that all S. equi strains had an identical sequence. For the S. zooepidemicus strains minor differences were found between the sodA sequences. The developed real-time PCR, based on the sodA and seeI genes was compared with conventional culturing on 103 cultured samples from horses with suspected strangles or other upper respiratory disease. The real-time PCR system was found to be more sensitive than conventional cultivation as two additional field isolates of S. equi and four of S. zooepidemicus were detected.
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Publication Date: 2007-04-08 PubMed ID: 17531409DOI: 10.1016/j.vetmic.2007.04.020Google Scholar: Lookup
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  • 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 researchers investigated the genetic characteristics of two subtypes of bacteria known to cause a contagious equine disease, using this to develop a more efficient detection method. They found that their new real-time PCR was more sensitive than traditional cultivation methods, allowing for better detection and differentiation of the bacteria.

Research Overview

  • The study focused on genetic characterization of clinical strains of two bacteria: Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus, both associated with a contagious equine disease called Strangles.
  • The goal was to design a real-time PCR (Polymerase chain Reaction) system capable of detecting and differentiating between the two types of bacteria, to improve diagnostic accuracy.

Methodology & Findings

  • The researchers performed gene-sequencing of 16S rRNA and sodA genes on 24 strains of each bacterium.
  • They found that while all S. equi strains had identical 16S rRNA sequences, the S. zooepidemicus strains could be divided into subgroups based on 16S rRNA sequencing. Interestingly, four strains, earlier identified biochemically as S. zooepidemicus, had sequences homologous with Streptococcus equi subsp. ruminatorum.
  • This hinted that classification of S. ruminatorum may not be solely determined by 16S rRNA sequencing. Moreover, the sodA gene sequence was identical among all S. equi strains, but showed minor differences among S. zooepidemicus strains.

Real-time PCR Development and Validation

  • The real-time PCR system was developed based on the sodA and seeI genes. It was tested against traditional cultivation methods using 103 cultured samples from horses suspected of Strangles or other upper respiratory diseases.
  • The new method showed greater sensitivity than conventional cultivation techniques, as it detected two additional field isolates of S. equi and four of S. zooepidemicus that were not identified by conventional methods.

Conclusions & Implications

  • The researchers concluded that their real-time PCR system is a more accurate and sensitive method for detecting and differentiating between S. equi and S. zooepidemicus.
  • Such enhancements in bacterial detection could lead to improved diagnosis and treatment of diseases like Strangles in horses.

Cite This Article

APA
Båverud V, Johansson SK, Aspan A. (2007). Real-time PCR for detection and differentiation of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus. Vet Microbiol, 124(3-4), 219-229. https://doi.org/10.1016/j.vetmic.2007.04.020

Publication

Veterinary microbiology
ISSN: 0378-1135
NlmUniqueID: 7705469
Country: Netherlands
Language: English
Volume: 124
Issue: 3-4
Pages: 219-229

Researcher Affiliations

Båverud, V
  • Department of Bacteriology, National Veterinary Institute, SE-751 89 Uppsala, Sweden. viveca.baverud@sva.se
Johansson, S K
    Aspan, A

      MeSH Terms

      • Animals
      • Base Sequence
      • DNA, Bacterial / chemistry
      • DNA, Bacterial / genetics
      • Genetic Variation
      • Horse Diseases / microbiology
      • Horses
      • Molecular Sequence Data
      • Phylogeny
      • Polymerase Chain Reaction / methods
      • Polymerase Chain Reaction / standards
      • Polymerase Chain Reaction / veterinary
      • RNA, Ribosomal, 16S / chemistry
      • RNA, Ribosomal, 16S / genetics
      • Respiratory Tract Infections / microbiology
      • Respiratory Tract Infections / veterinary
      • Sensitivity and Specificity
      • Sequence Analysis, DNA
      • Species Specificity
      • Streptococcal Infections / microbiology
      • Streptococcal Infections / veterinary
      • Streptococcus equi / classification
      • Streptococcus equi / genetics

      Citations

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