用于非洲猪瘟病毒多靶标核酸检测的DNA假病毒制备及定量

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中图分类号：S852.659.1 文献标志码：A 文章编号：0366-6964（2025）07-3555-06

Abstract： The study aims to develop a DNA pseudovirus with full quality control and avoiding plasmid cross-contamination in nucleic acid detection for African swine fever virus （ASFV）. The B646L/CD2v/MGF505-1R/MGF360-12L key-genes of ASFV and EGFP gene were co-combined into the baculovirus transfer vector pFastBacTMDual. The Bacmid pFastBac-EGFP-ASFV was formed by transformation. The Bacmid was transfected into sf21 cells and packaged to produce AcMNPV-EGFP-ASFV. The green fluorescence signal of EGFP protein expressed indicated that AcMNPV-EGFP-ASFV was preparated successfully. Genome traceability analysis of the AcMNPV-EGFP-ASFV showed that the ASFV target gene sequence was met the requirements of current detection methods. The ASFV quality control product is uniform and stable，and is stably stored at -20°C for at least 24 months. The quantification is 4. 09×103 copies. μL-1 （204号 using ddPCR. The preparation of ASFV DNA pseudovirus provides a new technical support for the monitoring of the African swine fever.

Keywords：African swine fever virus；DNA pseudovirus;quantification ∗ Corresponding author： WU Shaoqiang，E-mail： sqwu@sina. com

2018年8月，我国首次报道发生在沈阳的非洲猪瘟（Africanswine fever，ASF）疫情[1-3]，该毒株基因型为基因II型，同Georgia2007/1株同属一个进化分支[2]。（剩余7900字）