Key result
LNA-based multiplex RT-qPCR assay achieves 100% agreement with comparator for differentiating PRRSV lineages.
The developed LNA-based multiplex RT-qPCR assay provides a sensitive and specific tool for PRRSV surveillance.
Validates robust analytical performance of the LNA multiplex RT-qPCR assay; leaves open prospective clinical validation before routine diagnostic use.
= 0.99) and low detection limits of 13-26 copies per reaction at 95% detection probability. Robustness testing, including 2h bench exposure and a 3°C thermocycler shift, produced negligible Cq changes, while a 1,000-fold competitor challenge indicated minimal cross-reactivity. Intra- and inter-assay variability were low, qualitative agreement with the comparator was 100%, and reagents remained stable for 30 weeks and after up to 10 freeze-thaw cycles. This LNA-based multiplex RT-qPCR assay provides a sensitive, specific, and operationally convenient tool for PRRSV surveillance and control programs.
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Gyurján et al. (2026) studied Betaarterivirus (PRRSV) infection. LNA-based one-step multiplex RT-qPCR assay vs. Comparator assay was evaluated on Assay performance (detection limits, robustness, and qualitative agreement). An LNA-based multiplex RT-qPCR assay demonstrated low detection limits of 13-26 copies per reaction and 100% qualitative agreement with a comparator for differentiating PRRSV lineages.
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