Strangles Research Using Real-Time PCR

Veterinarians have to be cautioned that any qPCR-positive result for S. equi needs to be taken seriously and trigger biosecurity protocols aimed at reducing spread.
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Strangles in a Paint Horse

The objective this study was to investigate molecular surrogates for S. equi subspecies equi (S. equi) viability in biological samples from horses with strangles.

New research has been published in the Equine Veterinary Journal titled, "Use of quantitative real-time PCR to determine viability of Streptococcus equi subspecies equi in respiratory secretions from horses with strangles."

Background

In recent years, molecular approaches have been able to characterise the viability of equine upper respiratory tract pathogens using absolute molecular quantitation as well as detection of transcripts for virulence genes.

Objectives

The objective this study was to investigate molecular surrogates for S. equi subspecies equi (S. equi) viability in biological samples from horses with strangles.

Study design

Retrospective cross-sectional study.

Methods

S. equi culture-positive and culture-negative upper airway secretions were assessed by qPCR at the genomic (gDNA) and complimentary DNA (cDNA) level for various target genes (SeM, SEQ2190, eqbE and szpSe). Absolute quantitation was performed using standard curves and the results were expressed as number of S. equi target genes per μl of gDNA or cDNA. Additionally, the presence or absence of S. equi gene expression for the various target genes was assessed and compared to the culture results.

Results

While all 21 culture-positive samples tested S. equi qPCR-positive, up to 43.7% and 18.9% of 64 culture-negative samples tested qPCR-positive at the gDNA and cDNA level, respectively. Significant differences in absolute quantitation for S. equi at the gDNA level were found between culture-positive and culture-negative samples. When absolute quantitation of S. equi target genes at the gDNA level was assessed with presence or absence of transcripts, there was a significantly higher S. equi target gene number in samples with expression of transcripts compared to samples with no expression of transcripts.

Main Limitations

The lack of standardisation of samples collected in the field and the delay from sample collection to samples processing may have negatively affected the cultivability of S. equi and mRNA quality.

Conclusions

Molecular viability for S. equi can be investigated by determining absolute quantitation and/or by detecting mRNA for specific target genes. However, veterinarians have to be cautioned that any qPCR-positive result for S. equi needs to be taken seriously and trigger biosecurity protocols aimed at reducing spread.

The article can be accessed from the Wiley Online Library at http://onlinelibrary.wiley.com/wol1/doi/10.1111/evj.12809/abstract.

Authors

N. Pusterla, Department of Medicine and Epidemiology, School of Veterinary Medicine University of California, Davis; C. M. Leutenegger, IDEXX Laboratories Inc., West Sacramento; S. M. Barnum, Department of Medicine and Epidemiology, School of Veterinary Medicine University of California, Davis; B. A. Byrne, Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine University of California, Davis.