摘要
为建立检测副流感病毒5型(PIV5)的荧光定量RT-PCR方法,针对PIV5 L基因设计筛选出一对特异性引物和探针,对退火温度、引物浓度、探针浓度进行了优化,并进行了敏感性、特异性和重复性试验。结果显示:最适退火温度为54.7℃,最适引物浓度为0.5μmol/L,最适探针浓度为0.1μmol/L。检测灵敏度为10 copies、0.8 TCID50,特异性结果显示与BPIV3等22种常见病毒及原辅材料均无交叉,重复性试验显示不同模板浓度重复检测变异系数均小于1.5%,重复性良好。
To establish a fluorescence quantitative RT-PCR method for detection of Parainfluenza virus 5(PIV5),a pair of specific primers and probes were designed and screened for PIV5 L gene.The annealing temperature,primer concentration and probe concentration were optimized,and the sensitivity,specificity and repeatability tests were revaluated respectively.The results showed that the optimal annealing temperature was 54.7℃,the optimal primer concentration was 0.5μmol/L,and the optimal probe concentration was 0.1μmol/L.The detection sensitivity was 10 copies and 0.8 TCID50,the specificity results showed that there was no cross-over with 22 common viruses and raw materials such as BPIV3,and the repeatability test showed that the coefficient of variation of different template concentrations was less than 1.5%,and the repeatability was good.
作者
吴华伟
秦义娴
陈晓春
高金源
邓永
刘丹
苏佳
薛青红
陈延飞
WU Hua-wei;QIN Yi-xian;CHEN Xiao-chun;GAO Jin-yuan;DENG Yong;LIU Dan;SU Jia;XUE Qing-hong;CHEN Yan-fei(China Institute of Veterinary Drug Control,Beijing 100081,China)
出处
《中国兽药杂志》
2021年第9期1-7,共7页
Chinese Journal of Veterinary Drug
基金
中国兽医药品监察所所级课题(201801)。
