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Veterinary and animal science2026; 33; 100713; doi: 10.1016/j.vas.2026.100713

Evaluation of two point-of-care molecular diagnostic platforms for rapid detection of equine Hendra virus.

Abstract: Hendra virus (HeV) is a lethal zoonotic pathogen endemic to eastern Australia, posing significant risks to equine and human health. Rapid field detection of HeV enables timely intervention and outbreak management. This study evaluated candidate point-of-care (POC) molecular diagnostic platforms for HeV detection in equine samples: including a loop-mediated isothermal amplification (DARQ RT-LAMP) assay and real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR). Comparative analytical evaluation demonstrated that RT-qPCR exhibited superior analytical sensitivity relative to DARQ RT-LAMP, with a limit of detection of 1 copy/µL compared with 1,000 copies/µL, respectively. On this basis, DARQ RT-LAMP was not progressed beyond initial analytical evaluation due to insufficient sensitivity for the intended application. Bayesian Latent Class Model (BLCM) analysis estimated diagnostic sensitivity of 63.1% (95%PI 48.8-76.1%) and 80.4% (95%PI 67.6-90.2%) for RT-qPCR with HUDSON-prepared samples and extracted RNA, respectively, with identical specificity of 96.5% (95%PI 85.9-99.9%) for both sample types in virus transport medium. In 10% EDTA blood, diagnostic sensitivity was 71.3% (95%PI 54.2-85.1%) and 88.3% (95%PI 74.2-97.3%), respectively, with comparable specificity. Preliminary assessment of repeatability and reproducibility was promising (CVs <10%), although further field studies are required. These findings demonstrate that RT-qPCR, combined with rapid HUDSON sample preparation, provides a feasible molecular POC approach for preliminary rule-in or exclusion of HeV infection in horses while confirmatory laboratory testing is pending, supporting early risk management and reduced occupational exposure.
© 2026 The Authors.
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Publication Date: 2026-05-30 PubMed ID: 42291516PubMed Central: PMC13253132DOI: 10.1016/j.vas.2026.100713Google Scholar: Lookup
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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.

Research Summary

  • This study evaluated two rapid molecular diagnostic tests designed for use at the point of care (POC) to detect Hendra virus (HeV) infection in horses.
  • They compared a loop-mediated isothermal amplification assay (DARQ RT-LAMP) and a real-time reverse transcription quantitative PCR assay (RT-qPCR), assessing sensitivity, specificity, and feasibility for field application.

Background

  • Hendra virus (HeV) is a deadly virus native to eastern Australia that can infect horses and humans, posing significant health risks.
  • Rapid detection of HeV in horses is critical to initiate timely interventions and control potential outbreaks.
  • Point-of-care (POC) diagnostics are tests that can be performed quickly at the location of the animal without the need to send samples to specialized labs.

Diagnostic Platforms Evaluated

  • DARQ RT-LAMP Assay: A loop-mediated isothermal amplification method designed for rapid amplification of viral RNA without the need for thermal cycling.
  • RT-qPCR: A real-time reverse transcription quantitative polymerase chain reaction which is the gold standard molecular test for detecting viral RNA with high sensitivity.

Analytical Sensitivity Comparison

  • RT-qPCR Sensitivity: Could detect as few as 1 viral RNA copy per microliter, reflecting very high sensitivity.
  • DARQ RT-LAMP Sensitivity: Detected down to 1,000 copies per microliter, substantially less sensitive than RT-qPCR.
  • Due to insufficient sensitivity, the DARQ RT-LAMP platform was not advanced beyond initial testing phases.

Diagnostic Performance in Different Sample Types

  • Bayesian Latent Class Model (BLCM) Analysis: Used to estimate diagnostic sensitivity and specificity without relying on a perfect reference standard.
  • RT-qPCR with HUDSON Sample Prep (rapid preparation method):
    • Diagnostic sensitivity was estimated at 63.1% (with 95% probability interval 48.8-76.1%) using virus transport medium samples.
    • Specificity was very high at 96.5% (95% PI 85.9-99.9%).
  • RT-qPCR with Extracted RNA:
    • Higher diagnostic sensitivity of 80.4% (95% PI 67.6-90.2%) was observed.
    • Specificity remained at 96.5% (95% PI 85.9-99.9%).
  • 10% EDTA Blood Samples:
    • Sensitivity with HUDSON prep: 71.3% (95% PI 54.2-85.1%).
    • Sensitivity with extracted RNA: 88.3% (95% PI 74.2-97.3%).
    • Specificity was comparable to virus transport medium samples.

Repeatability and Reproducibility

  • Preliminary evaluation of test consistency showed coefficient of variation values under 10%, indicating good repeatability and reproducibility.
  • However, further field studies are necessary to confirm these findings in real-world conditions.

Implications and Conclusions

  • The RT-qPCR combined with rapid HUDSON sample preparation is a practical POC diagnostic approach to quickly rule in or exclude Hendra virus infection in horses.
  • This approach supports timely decision making and outbreak management by enabling preliminary diagnosis while awaiting confirmatory laboratory testing.
  • Early detection via POC testing can reduce occupational exposure risk to humans and enhance horse health management in Hendra virus endemic regions.
  • The DARQ RT-LAMP assay’s lower sensitivity limits its usefulness for this specific application.

Cite This Article

APA
Hulse L, Izzard L, Nagendrakumar SB, Colling A, Underwood D, Driver L, Williams DT, Ahern B. (2026). Evaluation of two point-of-care molecular diagnostic platforms for rapid detection of equine Hendra virus. Vet Anim Sci, 33, 100713. https://doi.org/10.1016/j.vas.2026.100713

Publication

Veterinary and animal science
ISSN: 2451-943X
NlmUniqueID: 101694897
Country: Netherlands
Language: English
Volume: 33
Pages: 100713
PII: 100713

Researcher Affiliations

Hulse, Lyndal
  • School of Veterinary Science, The University of Queensland, Gatton QLD 4343, Australia.
Izzard, Leonard
  • CSIRO, Australian Centre for Disease Preparedness, Geelong VIC 3220, Australia.
Nagendrakumar, Singanallur Balasubramanian
  • CSIRO, Australian Centre for Disease Preparedness, Geelong VIC 3220, Australia.
  • World Organization for Animal Health Collaborating Centre for Diagnostic Test Validation Science in the Asia-Pacific Region, CSIRO, Australian Centre for Disease Preparedness, Geelong VIC 3220, Australia.
Colling, Axel
  • CSIRO, Australian Centre for Disease Preparedness, Geelong VIC 3220, Australia.
  • World Organization for Animal Health Collaborating Centre for Diagnostic Test Validation Science in the Asia-Pacific Region, CSIRO, Australian Centre for Disease Preparedness, Geelong VIC 3220, Australia.
Underwood, Darren
  • Biosecurity Sciences Laboratory, Department of Primary Industries, Coopers Plains, QLD 4108, Australia.
Driver, Luke
  • Biosecurity Sciences Laboratory, Department of Primary Industries, Coopers Plains, QLD 4108, Australia.
Williams, David T
  • CSIRO, Australian Centre for Disease Preparedness, Geelong VIC 3220, Australia.
Ahern, Benjamin
  • School of Veterinary Science, The University of Queensland, Gatton QLD 4343, Australia.

Conflict of Interest Statement

The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Dr Lyndal Hulse reports financial support was provided by AgriFutures. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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