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Detection of Streptococcus equi subsp. equi in guttural pouch lavage samples using a loop-mediated isothermal nucleic
Ashley G Boyle1, Shelley C Rankin2, Kathleen O'Shea2
1Department of Clinical Studies-New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, Pennsylvania, USA.
Journal of Veterinary Internal Medicine
|March 17, 2021
Summary
A new LAMP assay for Streptococcus equi subsp. equi (S. equi) shows promise for rapid point-of-care strangles detection. The microfluidic device offers comparable accuracy to qPCR, aiding in early identification and disease control.
Area of Science:
- Veterinary Diagnostics
- Microbiology
- Molecular Biology
Background:
- Strangles, caused by Streptococcus equi subsp. equi (S. equi), poses a significant threat to equine health.
- Rapid point-of-care (POC) diagnostics are crucial for controlling strangles by identifying infected and carrier horses.
Purpose of the Study:
- To evaluate the diagnostic performance of an eqbE isothermal amplification (LAMP) assay for S. equi detection.
- To compare the eqbE LAMP assay, in both benchtop and microfluidic formats, against a standard triplex real-time quantitative polymerase chain reaction (qPCR) assay.
Main Methods:
- Sixty-eight guttural pouch lavage (GPL) specimens from horses recovering from strangles were retrospectively tested.
- The eqbE LAMP assay (benchtop and microfluidic) and a triplex qPCR assay targeting eqbE and SEQ2190 genes were performed.
- Diagnostic accuracy was assessed using receiver operating characteristic (ROC) analysis with the triplex qPCR as the reference standard.
Main Results:
- The benchtop eqbE LAMP detected S. equi in 27/68 specimens, and the microfluidic device in 31/64.
- Both LAMP formats demonstrated excellent diagnostic discrimination, with ROC areas of 0.813 (benchtop) and 0.811 (microfluidic).
- No significant difference in performance was observed between the benchtop and microfluidic LAMP assays (P = .97).
Conclusions:
- The eqbE LAMP microfluidic device accurately detects S. equi in equine GPL specimens.
- This assay holds potential for development into a rapid, sensitive, accurate, and cost-effective POC diagnostic tool for strangles.
- Early detection via POC testing can significantly aid in managing and reducing strangles transmission in horse populations.
