激光生物学报2023，Vol.32Issue(6)：554-560,7.DOI:10.3969/j.issn.1007-7146.2023.06.008
非洲猪瘟病毒B646L和F778R双基因PCR检测方法的构建
Construction of PCR Detection Method for African Swine Fever B646L and F778R Genes
摘要
Abstract
In order to detect African swine fever virus(ASFV)quickly by ordinary PCR,In this study,we established a com-mon PCR method for ASFV B646L and F778R dual genes by two pairs of specific primers which were designed based on the target sequences of ASFV B646L and F778R genes respectively.And its specificity,sensitivity and reproducibility were tested.It showed that the optimal combination of PCR system(25.0 μL system)was Pfu enzyme 0.2 μL,10×Reaction Buffer 2.5 μL,2.5 mmol/L dNTP Mixture 2.5 μL,B646L upstream primer and B646L downstream primer were 1.5 μL,F778R upstream primer,F778R downstream primer each 0.5 μL,DNA template 1.0 μL,annealing temperature 57℃,annealing time 30 s.The method was only used for specific amplification of ASFV B646L and F778R genes,without cross-reaction.The lower limits of detection of positive plasmid B646L and F778R were 7.2×107 copies/μL and 1.5×106 copies/μL.The test repeatability is good.This study has important value for improving the accuracy,specificity and efficiency of clinical diagnosis of ASFV,and provides reliable technical support for rapid detection of ASFV.关键词
非洲猪瘟病毒/多重PCR/B646L基因/F778R基因/快速检测Key words
African swine fever/multiplex PCR/B646L gene/F778R gene/quick detection分类
农业科技引用本文复制引用
王慧,钟菊花,杨俊,杜丽飞,王红兵,刘俊琦..非洲猪瘟病毒B646L和F778R双基因PCR检测方法的构建[J].激光生物学报,2023,32(6):554-560,7.基金项目
湖南现代农业产业技术体系项目(湘[2022]222号). （湘[2022]222号）
