检测鱼传染性造血器官坏死病毒重组酶聚合酶扩增方法的建立及初步应用  

Establishment and Preliminary Application of Recombinase Polymerase Amplification Method for Detecting Infectious Hematopoietic Necrosis Virus

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)

机构地区：[1]烟台海关技术中心,山东烟台264000 [2]上海医药集团青岛国风药业股份有限公司,山东青岛266000

出　　处：《动物医学进展》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)

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.

关 键 词：重组酶聚合酶扩增技术 快速检测 鱼类传染性造血器官坏死病毒 

分 类 号：S854.43[农业科学—临床兽医学] S941.414[农业科学—兽医学]