Key result
The triplex real-time PCR assay simultaneously detected PRV, PTV1, and SS2 with a detection limit of 1.0 × 10^0 copies/μL and showed complete concordance with commercial singleplex kits.
Why the study?
Overlapping clinical presentations and pathological alterations among PRV, PTV1, and SS2 complicate clinical differentiation of porcine neurological disorders, necessitating a sensitive and specific diagnostic method for concurrent detection.
The developed triplex real-time PCR assay provides a highly sensitive and specific diagnostic tool for the simultaneous detection of PRV, PTV1, and SS2 in swine populations.
Answers are grounded in this paper and cite their sources.
May aid simultaneous diagnosis of overlapping porcine neurologic pathogens; leaves open broader field validation.
Introduction Porcine neurological disorders represent a prevalent clinical condition that leads to significant mortality and economic losses within the swine industry. Pseudorabies virus (PRV), porcine teschovirus 1 (PTV1), and Streptococcus suis 2 (SS2) are key viral and bacterial pathogens implicated in the manifestation of neurological symptoms in pig populations. The overlapping clinical presentations and pathological alterations associated with these pathogens pose challenges in their clinical differentiation. Therefore, it is essential to develop a diagnostic method with high sensitivity and specificity that can simultaneously detect and differentiate these viral and bacterial agents. Materials and methods A triplex real-time PCR assay using TaqMan probes was developed to simultaneously detect PRV, PTV1, and SS2. To assess the efficacy of the established assay, 30 clinical samples of animals with nervous symptoms were used to compare the results obtained from the triplex real-time PCR assay with those obtained from commercial singleplex real-time PCR kits. Furthermore, a total of 282 samples were tested and analyzed to validate the utility of the assay. Results The triplex real-time PCR assay exhibited high sensitivity, specificity, and repeatability, with a detection limit of 1.0 × 10 0 copy/μL. The triplex real-time PCR method and commercial singleplex real-time PCR kits showed complete concordance in detecting PRV, PTV1, and SS2. Clinical data indicated single infection rates of 8.16% for PRV, 26.95% for PTV1, and 7.80% for SS2. The observed co-infection rates were 7.45% for PRV + PTV1, 0.71% for PRV + SS2, 1.42% for PTV1 + SS2, and 1.77% for PRV + PTV1 + SS2, respectively. Conclusion The triplex real-time PCR method developed in this study effectively distinguishes PRV, PTV1, and SS2 simultaneously, serving as a valuable diagnostic tool. This method is anticipated to play a crucial role in preventing and controlling infectious disease spread and supporting epidemiological investigations.
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Lai et al. (2025) studied Porcine neurological disorders (n=282). TaqMan triplex real-time PCR assay vs. Commercial singleplex real-time PCR kits was evaluated on Limit of detection and diagnostic concordance. The triplex real-time PCR assay simultaneously detected PRV, PTV1, and SS2 with a detection limit of 1.0 × 10^0 copies/μL and showed complete concordance with commercial singleplex kits.
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