Real-time fluorometric and end-point colorimetric isothermal assays for detection of equine pathogens C. psittaci and equine herpes virus 1: validation, comparison and application at the point of care.
Authors: Jelocnik Martina, Nyari Sharon, Anstey Susan, Playford Nicole, Fraser Tamieka A, Mitchell Keith, Blishen Anna, Pollak Nina M, Carrick Joan, Chicken Catherine, Jenkins Cheryl
Journal: BMC veterinary research
Summary
# Editorial Summary The emergence of *Chlamydia psittaci* as a reproductive pathogen in Australian Thoroughbreds, alongside the established threat of equine herpesvirus-1 (EHV-1), has highlighted the need for rapid diagnostic tools beyond traditional laboratory-based molecular assays. Jelocnik and colleagues developed and validated loop-mediated isothermal amplification (LAMP) assays for both pathogens, comparing three detection formats: real-time fluorometric (rtLAMP), colorimetric (cLAMP), and SYBR Green end-point detection (sgLAMP) against reference PCR methods using clinical field samples. The LAMP assays demonstrated reliable sensitivity and specificity comparable to standard diagnostics, with the critical advantage that colorimetric and SYBR Green formats could be read with the naked eye without specialised equipment—a feature that proved feasible during two pilot point-of-care studies conducted in an equine hospital setting. For practitioners managing reproductive disease outbreaks, these findings suggest that isothermal amplification assays offer a viable pathway towards decentralised diagnostics, potentially enabling faster decision-making on isolation protocols and treatment without the delays inherent to sending samples to distant laboratories. The practical validation in real clinical environments strengthens the case for integrating LAMP assays into equine reproductive disease surveillance, though further work establishing cost-effectiveness and uptake barriers across different practice settings would be valuable for widespread implementation.
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Practical Takeaways
- •LAMP-based testing offers a viable alternative to laboratory PCR for rapid on-site diagnosis of C. psittaci and EHV-1 in equine abortion cases
- •Colorimetric readouts eliminate need for specialized equipment, making this technology accessible for equine hospitals and field settings
- •Earlier diagnosis of abortigenic pathogens enables faster isolation protocols and reduces transmission risk in breeding operations
Key Findings
- •Real-time LAMP assays for C. psittaci and EHV-1 were successfully validated against reference diagnostic assays
- •Colorimetric LAMP (cLAMP) and SYBR Green LAMP (sgLAMP) enabled naked-eye end-point detection suitable for point-of-care testing
- •Point-of-care LAMP assays were successfully applied in pilot studies at an equine hospital, demonstrating clinical feasibility