Study And Application Of Cross-primers Isothermal Amplification Methods For Detection Of Five Soybean Seedbrone Plant Quarantine Virus

China is the world’s first soybean importing country,soybean inspection and quarantine is particularly important.Southern bean mosaic virus（SBMV）,Tomato ringspot virus（To RSV）,Arabis mosaic virus（Ar MV）,Tobacco ringspot virus（TRSV）and Bean pod mottle virus（BPMV）are soybean seedborne plant quarantine viruses that are frequently intercepted at ports and customs.With the rapid development of domestic and international trade,the import of soybean in China continues to increase,and the risk of soybean seed-borne phytosanitary virus entering China is constantly increasing.Therefore,we developed the rapid and accurate detection method for the five soybean seedborne plant quarantine viruses by using cross-primer isothermal amplification（CPA）technology,and the results are as follows.According to the coat protein gene sequences of SBMV,To RSV,Ar MV,TRSV and BPMV,specific primers were designed and screened respectively,and a one-step cross-primer isothermal amplification technique for detection of five viruses was established.The method could be used for single detection of SBMV,To RSV,Ar MV,TRSV and BPMV,respectively.It has good specificity.The sensitivity of the method combined with nucleic acid strip for detection of SBMV was 5 ng/μl total RNA,and the sensitivity of the method combined with real-time fluorescence detection of SBMV was 5×10^-3ng/μl total RNA.Combined with nucleic acid strip,the sensitivity of To RSV,Ar MV,TRSV and BPMV was 0.5 ng/μL of total RNA.The sensitivity of To RSV,Ar MV,TRSV and BPMV was 0.5 ng/μL,0.05ng/μL,5ng/μL and 0.05ng/μL,respectively.