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A method for the analysis of African swine fever by viral metagenomic sequencing
  • +12
  • ChiHai Ji,
  • JingZhe Jiang,
  • Yingfang Wei,
  • ZhiYuan Wang,
  • Yongjie Chen,
  • ZhanZhuo Mai,
  • MengKai Cai,
  • ChenXiao Qin,
  • Yu Cai,
  • HeYou Yi,
  • Guan Liang,
  • Chihai Ji,
  • Lang Gong,
  • Guihong Zhang,
  • Heng Wang
ChiHai Ji
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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JingZhe Jiang
South China Sea Fisheries Research Institute
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Yingfang Wei
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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ZhiYuan Wang
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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Yongjie Chen
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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ZhanZhuo Mai
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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MengKai Cai
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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ChenXiao Qin
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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Yu Cai
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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HeYou Yi
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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Guan Liang
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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Chihai Ji
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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Lang Gong
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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Guihong Zhang
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases
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Heng Wang
Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases

Corresponding Author:[email protected]

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Abstract

In 2018, there was an outbreak of African swine fever (ASF) in China, which spread to other provinces in the following three years, severely damaging China’s pig industry. ASF is caused by the African swine fever virus (ASFV). Given that the genome of the African swine fever virus is very complex and whole genome information is currently inadequate, it is important to efficiently obtain virus genome sequences for genomic and epidemiological studies. The prevalent ASFV strains have low genetic variability; therefore, whole genome sequencing analysis provides a basis for the study of ASFV. We provide a method for the efficient sequencing of whole genomes, requiring only a small number of tissues. The pig spleen were crushed and homogenized, and the tissue and cell fragments were removed by low-speed centrifugation. The virus-like particles (VLPs) in the solution were purified and concentrated by filtration and over-speed centrifugation. After obtaining the relatively pure VLP sample, nucleic acids were extracted simultaneously by co-extraction of viral DNA (dsDNA, ssDNA) and RNA (ssRNA, dsRNA). Then, the database construction method was selected according to the genomic types of ASFV, and the whole ASFV genome was obtained through data filtering, host sequence removal, virus classification, data assembly, virus sequence identification, statistical analysis, gene prediction, and functional analysis. Our proposed method will facilitate ASFV genome sequencing and novel virus discovery.