摘要
为建立鱼类传染性造血器官坏死病毒(IHNV)的快速检测方法,根据IHNV的G基因保守序列设计特异性引物,建立了基于重组酶聚合酶扩增(RPA)的IHNV检测方法。该方法在30℃20min即可完成,对IHNV具有良好的特异性和敏感性,最低检出限为156ng/mL;用建立的RPA和RT-PCR对实验室保存的50份样品进行检测,结果显示,RPA的阳性检出率为14.00%,RT-PCR的阳性检出率为12.00%,说明RPA比RT-PCR具有更高的灵敏度。建立的RPA方法为IHNV的实验室检测提供了更多选择。
A rapid dection method of Infectious hematopoietic necrosis virus(IHNV)based on the recombinase polymerase amplification was established using primers designed according to G gene.The method could be finished in 20 min with good specificity and sensitivity.The optimal amplification temperature of the method was 30℃and the detection limit was 156 ng/mL.Moreover,a total of 50 laboratory-preserved samples were detected by RPA and RT-PCR,respectively.The results showed that the positive rate of RPA was 14.00%,and that of PCR was 12.00%.The positive detection rate of RPA was slightly higher than that of RT-PCR,which indicated that RPA had higher sensitivity than RT-PCR.The quick,simple,specific and sensitive method without precision temperature control instrument was suitable for common laboratory.
作者
梁君妮
尹伟力
刘鹏
马晓玲
段效辉
林森
LIANG Jun-ni;YIN Wei-li;LIU Peng;MA Xiao-ling;DUAN Xiao-hui;LIN Sen(Technology Center of Yantai Customs,Yantai,Shandong,264000,China;Shanghai Pharmaceutical Group Qingdao Guofeng Pharmaceutical Co.LTD,Qingdao,Shandong,266000,China)
出处
《动物医学进展》
北大核心
2020年第2期14-17,共4页
Progress In Veterinary Medicine
基金
国家重点研发计划项目(2017YFF0211103)
国家质量监督检验检疫总局科研项目(2017IK232)
