Cross-priming amplification-based lateral flow strip as a novel tool for rapid on-site detection of wild-type pseudorabies virus
Abstract Pseudorabies (PR), also called as Aujeszky’s disease, is an important infectious disease of pigs and other mammals caused by pseudorabies virus (PRV). This paper presents a novel assay based on cross-priming amplification in combination with gold nanoparticle lateral flow strip (CAP-strip) for rapid and on-site detection of wild-type PRV. In the assay, FITC and biotin were labeled at the 5′-ends of the two probes, respectively, resulting in amplified products labeled with either FITC or biotin at the 5′-end. Then the amplified products (biotin-dsDNA-FITC), as novel targets, were recognized by standardized strips. The assay had excellent visualization results from 2.0 × 10 2 to 2.0 × 10 8 copies of PRV and the detection limit was determined to be 200 copies. No cross-reactivity was observed with several common porcine viruses. When testing a total of 293 field swine samples, the agreement between the CPA-strip assay and a triplex real-time PCR or virus isolation were 100% (293/293) or 97.3% (285/293). Collectively, the assay is specific, sensitive and convenient that enables rapid, on-site diagnosis of PRV infections. Highlights Cross-priming amplification-based lateral-flow strip was developed for detection of wild-type pseudorabies virus. The assay combined the sensitivity of cross-priming amplification with the convenience of the strip. The labeled amplicons were detected by standardized strips. The assay was able to detect as low as 200 copies of the virus. The assay can be used for rapid, on-site diagnosis of pseudorabies.
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