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

doi:10.11843/j.issn.0366-6964.2025.07.047

Abstract

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 pFastBac^TMDual. 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 ℃ for at least 24 months. The quantification is 4.09×10³ copies·μL^-1 using ddPCR. The preparation of ASFV DNA pseudovirus provides a new technical support for the monitoring of the African swine fever.

Key words: African swine fever virus, DNA pseudovirus, quantification

CLC Number:

S852.659.1

DENG Junhua, LI Haoxuan, CHEN Dongjie, LV Jizhou, WANG Jingjing, ZHANG Zhou, YUAN Xiangfen, WEI Fang, WU Shaoqiang. Preparation and Quantification of DNA Pseudovirus with Multi-Target Nucleic Acid Detection for African Swine Fever Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(7): 3555-3560.

Figures/Tables 3

References 13

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Preparation and Quantification of DNA Pseudovirus with Multi-Target Nucleic Acid Detection for African Swine Fever Virus

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