Application and Building of Immunocapture Real-time RT-PCR for Detection of Cucumber Mosaic Virus in Individual Peach Aphid (Myzus persicae)

SONG Liyun1,YANG Jinguang1,LI Xihong2,SONG Yuchuan3,ZHANG Changhua4,ZHAN Huaixu1,SHEN Lili1,QIAN Yumei1,WANG Fenglong1(1.Open Project Program of Key Laboratory of Tobacco Pest Monitoring Controlling & Integrated Management,State Tobacco Monopoly Bureau,Qingdao 266101,China;2.Tobacco Research Institute of Hubei Province,Wuhan 430030,China;3.Baoshan Oriental Tobacco Company,Baoshan,Yunnan 678000,China;4.Zunyi Company of Guizhou Provincial Tobacco Corporation,Zunyi,Guizhou 563000,China)

immunocapture real-time RT-PCR;Cucumber mosaic virus(CMV);myzus persicae

Cucumber mosaic virus(CMV)is transmitted in nonpersistent manner by peach aphid(Myzus persicae),leading to many plant diseases.The large number of viruliferous peach M.persicae is one of the major culprits for the rampant outbreak of CMV disease in tobacco.In present study,Immunocapture real-time RT-PCR(IC-real-time RT-PCR) was investigated to detect CMV in individual peach aphid(M.persicae) by combining Enzyme-Linked Immunosorbent Assay(ELISA) and conventional RT-PCR.The complex of CMV CP antibodies and CMV particles was adhered to PCR tubes wall,then reverse transcriptase reaction was conducted directly,subsequently real-time PCR.Compared with conventional RT-PCR and ELISA to detect CMV in individual peach aphid,this method was more sensitive and specific.This assay of RT-PCR could be directly carried out without total RNA extraction by capturing CMV particle through CMV CP antibodies,and effectively reduce the pollution and inhibiting effects of proteins,polysaccharide and phenols materials in operating process.Therefore,as a specific,rapid,sensitivity and simple method for detecting slight amount CMV particle,IC-RT-PCR has significance of prediction,forecast and prevention of tobacco virus diseases.