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\begin{document}

\title{Establishment and validation of a duplex, real-time PCR assay for
detection of African swine fever virus}



\author[1]{Yang Zhan}%
\author[2]{Luhua Zhang}%
\author[3]{Yuan  Lin}%
\author[1]{Yunfeng Cai}%
\author[2]{Yawen Zou}%
\author[4]{Zhiyou Hao}%
\author[5]{Zhanhua Luo}%
\author[2]{Naidong Wang}%
\author[1]{Zhibang Deng}%
\author[1]{Yi Yang}%
\author[3]{Changjian Wang}%
\author[1]{Qing Yang}%
\affil[1]{Hunan Agricultural University}%
\affil[2]{Affiliation not available}%
\affil[3]{Animal Disease Prevention and Control Center of Hunan Province}%
\affil[4]{Yongzhou Animal Husbandry and Aquatic Affairs Center}%
\affil[5]{Dingcheng Animal Husbandry and Aquatic Affairs Center}%


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  \date{June 17, 2024}


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\begin{abstract}
The outbreak of African swine fever (ASF) in China in 2018, caused a
huge economic loss on the pig industry. To accurately diagnose clinical
infection with ASF virus (ASFV), we developed a duplex
TaqMan-probe-based real-time PCR for the simultaneous detection of two
discontinuous genes in the virus genome to avoid false-negative in the
clinical diagnosis. Two sets of ASFV-gene specific primers, along with
two TaqMan fluorescent probes were designed to target the conserved
region of B646L and B438L genes, respectively. The method showed high
sensitivity and specificity, with a detection limit of 10 copies/\selectlanguage{greek}μ\selectlanguage{english}l
without cross-reactions with other pathogen genomes of CSFV, PRRSV,
PEDV, PRV, PPV and PCV2. 269 clinical swine samples were tested using
this method, and the results of 60 samples showed excellent consistency
(95.0\%) with a commercial kit. This method will greatly improve the
efficiency for surveillance of ASFV and can help reduce economic losses
to the swine industry, which also benefits animal and public health.%
\end{abstract}%



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