Correction: Villalba et al. Development and Validation of Three Triplex Real-Time RT-PCR Assays for Typing African Horse Sickness Virus: Utility for Disease Control and Other Laboratory Applications. Viruses 2024, 16, 470.

Correction: Villalba et al. Development and Validation of Three Triplex Real-Time RT-PCR Assays for Typing African Horse Sickness Virus: Utility for Disease Control and Other Laboratory Applications. Viruses 2024, 16, 470.

In the original publication […].

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Overview

• This research article describes the development and validation of three triplex real-time RT-PCR assays designed to accurately identify and type African Horse Sickness Virus (AHSV) strains.
• The assays have practical applications for disease control and laboratory diagnostics, enhancing the ability to quickly and reliably diagnose AHSV infections.

Introduction

• African Horse Sickness Virus (AHSV) is a significant pathogen affecting equids, characterized by high mortality rates and severe economic impact.
• AHSV comprises multiple serotypes, and rapid, accurate typing of these serotypes is critical for outbreak management and vaccine strategy.
• Current diagnostic methods have limitations in sensitivity, specificity, or speed.
• The study addresses the need for improved molecular diagnostic tools that allow simultaneous detection and differentiation of AHSV serotypes.

Methodology

• Three triplex real-time reverse transcription PCR (RT-PCR) assays were designed, each capable of detecting three different AHSV serotypes simultaneously.
• Primers and probes were carefully designed to target conserved yet type-specific genetic regions of the AHSV genome.
• Assays were optimized and validated using a comprehensive panel of reference strains and clinical samples.
• Analytical sensitivity (limit of detection), specificity (cross-reactivity with related orbiviruses), and reproducibility (intra- and inter-assay variation) were assessed rigorously.

Results

• The triplex assays demonstrated high sensitivity, detecting viral RNA at low copy numbers.
• They showed excellent specificity, with no cross-reactivity observed with other orbiviruses or non-target serotypes.
• The tests reliably differentiated among the different AHSV serotypes included in each triplex assay.
• The assays were robust and reproducible, suitable for high-throughput diagnostic laboratories.
• Application to field and experimental samples confirmed their practical utility in real-world settings.

Discussion and Implications

• The newly developed triplex real-time RT-PCR assays provide a rapid and accurate method for typing AHSV, which is essential for timely disease control measures such as quarantine and vaccination.
• The assays enable simultaneous detection of multiple serotypes in a single reaction, improving laboratory efficiency and reducing costs.
• These tools enhance epidemiological surveillance, outbreak investigation, and vaccine matching.
• The approach could be adapted for other multi-serotype viral diseases where rapid typing is crucial.

Conclusion

• The study successfully developed and validated three triplex real-time RT-PCR assays for African Horse Sickness Virus typing.
• The assays are suitable for routine laboratory use, supporting improved disease control strategies and research applications.
• This advancement contributes significantly to equine health management and AHSV outbreak mitigation globally.