Key result
New multiplex RT-PCR assay detects ASFV in ~44% of suspected infected piglets alongside CSFV and APPV.
Why the study?
African swine fever virus, classical swine fever virus, and atypical porcine pestivirus cause considerable losses to the pig industry, making early and accurate simultaneous diagnosis critical.
Cross-Sectional (n=384)
The developed mRT-PCR assay provides a highly specific and sensitive tool for the rapid simultaneous detection of ASFV, CSFV, and APPV in swine.
May enable simultaneous swine virus detection in outbreaks; leaves open field validation before routine diagnostic adoption.
African swine fever virus (ASFV), classical swine fever virus (CSFV) and atypical porcine pestivirus (APPV) have caused considerable financial losses to the pig industry worldwide, and it is critical to achieve early and accurate diagnosis of these viruses to control the diseases induced by them. In this study, three pairs of specific primers were designed based on the highly conserved genome regions of these viruses, and a multiplex reverse transcription-polymerase chain reaction (mRT-PCR) assay for ASFV, CSFV and APPV was established after various reaction conditions were optimized. The mRT-PCR assay consisted of two steps, that is, reverse transcription (RT) and mPCR. The assay was highly specific, sensitive, and reproducible for ASFV, CSFV and APPV without cross-reaction with other swine pathogens. The sensitivity of this assay, which used purified plasmid constructs containing specific viral target fragments as templates, was 6.34 × 102 copies/μL for ASFV and 6.34 × 101 copies/μL for both CSFV and APPV. A total of 384 clinical samples from piglets suspected to be infected in Guangxi Province, Southern China, during 2018−2019 were analyzed by the established mRT-PCR method. The results showed that the positive rates of ASFV, CSFV and APPV were 43.75 %, 13.28 % and 4.17 %, respectively, and the coinfection rates of ASFV/CSFV, ASFV/APPV and CSFV/APPV were 5.47 %, 1.83 % and 1.30 %, respectively. To understand the epidemiological characteristics of APPV, the newly discovered virus, in Guangxi Province, the clinical samples from APPV-positive animals were selected randomly for amplification and sequencing, and the complete genomic sequences of four APPV strains were obtained. Phylogenetic analysis demonstrated that APPV strains from Guangxi Province had a high degree of genetic diversity. This study provides an important tool for rapid detection and accurate diagnosis of ASFV, CSFV and APPV.
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Liu et al. (2020) conducted a cross-sectional in African swine fever virus (ASFV), classical swine fever virus (CSFV) and atypical porcine pestivirus (APPV) infection (n=384). Multiplex RT-PCR assay was evaluated on Positive rates of ASFV, CSFV, and APPV. A newly developed multiplex RT-PCR assay detected positive rates of 43.75%, 13.28%, and 4.17% for ASFV, CSFV, and APPV, respectively, in 384 clinical samples from suspected infected piglets.
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