期刊论文

中文

激光生物学报

1007-7146

2023

32

6

554-560

10.3969/j.issn.1007-7146.2023.06.008

:湖南现代农业产业技术体系项目

本校为其他机构

为建立一种能够快速检测非洲猪瘟病毒(ASFV)的普通PCR方法, 本研究分别以ASFV B646L和F778R基因为靶序列设计了两对特异性引物, 建立ASFV B646L和F778R双基因普通PCR方法, 并对其特异性、灵敏性及重复性进行检测。结果显示: 该PCR体系(25.0 μL体系)的最优组合为Pfu酶0.2 μL、10×Reaction Buffer 2.5 μL、2.5?mmol/L dNTP Mixture 2.5 μL、B646L上下游引物各1.5 μL、F778R上下游引物各0.5 μL、DNA模板1.0 μL; 退火温度为57℃、退火时间为30 s、72℃延伸2.5 min, 30个循环。该方法仅针对ASFV B646L和F778R基因进行特异性扩增, 无交叉反应。同时对B646L和F778R阳性质粒的最低检测下限分别为...

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 prime...