基于PCR-LFIA的非洲猪瘟病毒核酸检测方法研究OA北大核心CSTPCD
Study on Nucleic Acid Detection Method of African Swine Fever Vi-rus Based on PCR-LFIA
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1.6×10-2～1.6×108 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 1.6×10-2-1.6×108 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.
向四意;苏晓娜;张咏仪;陈俊杰;杨旭琼;沈玉栋;杨金易
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
化学
非洲猪瘟聚合酶链式反应(PCR)免疫层析可视化
African swine feverpolymerase chain reaction(PCR)immunochromatographyvi-sualization
《分析测试学报》 2024 (006)
928-932 / 5
新型生物快速检测技术研究及应用开发(h20230751)
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