基于PCR-LFIA的非洲猪瘟病毒核酸检测方法研究  

Study on Nucleic Acid Detection Method of African Swine Fever Vi⁃rus Based on PCR-LFIA

作　　者：向四意 苏晓娜 张咏仪 陈俊杰 杨旭琼 沈玉栋[1] 杨金易[1] XIANG Si-yi;SU Xiao-na;ZHANG Yong-yi;CHEN Jun-jie;YANG Xu-qiong;SHEN Yu-dong;YANG Jin-yi(Guangdong Provincial Key Laboratory of Food Quality and Safety,College of Food Science,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学食品学院广东省食品质量安全重点实验室,广东广州510642

出　　处：《分析测试学报》2024年第6期928-932,共5页Journal of Instrumental Analysis

基　　金：新型生物快速检测技术研究及应用开发(h20230751)。

摘　　要：该文利用聚合酶链式反应(PCR)技术进行扩增后,以侧流免疫层析方法(LFIA)对核酸进行检测,建立了一种非洲猪瘟病毒核酸的快速检测方法。通过设计引物,构建了pUC57-ASFV-(1-1941)质粒作为阳性质控品,采用柠檬酸三钠还原法制备胶体金颗粒用于标记抗体建立免疫分析方法。通过优化实验条件,在质粒浓度1.6×10^(-2)~1.6×10^(8)copies/μL范围内,得到pUC57-ASFV-(1-1941)阳性质粒的检出限为1.6 copies/μL。该方法与其他猪病毒无交叉,特异性良好,可作为非洲猪瘟现场筛查的辅助方法。In this paper,polymerase chain reaction(PCR)was used to amplify nucleic acid,lateral flow immunoassay(LFIA)was used to detect nucleic acid,and a rapid detection method for African swine fever virus nucleic acid was established.In this study,by designing primers,the positive plasmid pUC57-ASFV-(1-1941)was constructed as a quality control product and colloidal gold par⁃ticles were prepared by trisodium citrate reduction method for antibody labeling to establish an immu⁃noassay method.Under the optimal experimental conditions,the plasmid concentration was in the range of 1.6×10^(-2)-1.6×10^(8)copies/μL,the detection limit of pUC57-ASFV-(1-1941)positive plas⁃mid was 1.6 copies/μL,the method had no cross with other pig viruses and had good specificity.The established method can be used as an auxiliary method for field screening of African swine fever.

关 键 词：非洲猪瘟 聚合酶链式反应(PCR) 免疫层析 可视化 

分 类 号：O657.3[理学—分析化学] R446.6[理学—化学]