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

## TL;DR

A new rapid diagnostic test for peste des petits ruminants (PPR) was developed, offering high accuracy and ease of use in various conditions.

## Contribution

A novel multienzyme isothermal and lateral flow dipstick assay targeting the N gene of PPRV was developed and validated.

## Key findings

- The assay detects PPRV with high sensitivity (10 copies/μL) and no cross-reactivity with other viruses.
- It provides consistent results with a coefficient of variation below 3.0% and a 100% positive detection rate in clinical samples.
- The test can be completed within 30 minutes at 42°C without requiring a laboratory setting.

## Abstract

In this study, we established a visual rapid diagnostic method for peste des petits ruminants and validated its sensitivity, specificity, and repeatability. The results demonstrate that this method can rapidly and accurately detect the peste des petits ruminants virus with sensitivity comparable to current mainstream detection methods. Moreover, it requires less operational time, demands lower expertise from operators, and can be performed even under extreme conditions without the need for a laboratory environment. The method shows no cross-reactivity with other common animal diseases, exhibits good repeatability, and produces consistent and highly accurate results for clinical samples compared to other established methods. Additionally, the detection outcomes can be presented in multiple visual formats and are easy to interpret.

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).