Rapid Detection of Peste Des Petits Ruminants via Multienzyme Isothermal and Lateral Flow Dipstick Combination Assay Based on N Gene

In this study, a multienzyme isothermal and lateral flow dipstick combination assay for PPRV detection was established, the designed primers and probes targeting the N gene were screened and optimized, and analytical sensitivity, specificity, and repeatability of developed method were systematically evaluated. The experimental results demonstrated that this method is easy to operate, can complete detection within 30 min at 42 °C, and is capable of detecting all lineages of peste des petits ruminants virus (PPRV) without cross-reactivity with other viruses. The limit of detection could reach 10 copies/μL. Repeatability validation showed that the coefficients of variation (CV) for both intra-assay and inter-assay experiments were below 3.0%. The positive detection rate for clinical samples could reach 100%. The test results are visually interpretable via fluorescence and lateral flow strips. In conclusion, this method exhibits high analytical sensitivity, specificity, and excellent repeatability, enabling rapid diagnosis of peste des petits ruminants (PPR).