甘薯羽状斑驳病毒(SPFMV)ELISA鉴定及RT-PCR检测方法的建立

ELISA Identification and Development of RT-PCR Detection of Sweet Potato Feathery Mottle Virus

利用双抗体酶联免疫吸附测定法（DAS-ELISA ）对采自福建省51份甘薯叶片进行检测，结果显示51份样品中有24份SPFM V呈阳性，阳性率为47.06％，其中泉州样品SPFM V阳性率最高，为71.40％，其次为南安和龙岩的样品，阳性率均为50％。根据GenBank中公布的SPFM V外壳蛋白（CP）基因序列保守区域设计了1对特异性引物，通过RT-PCR反应程序的优化建立了能检测SPFM V的检测方法。该检测方法能够扩增出SPFM V特异性片段，片段大小为441 bp ，测序结果表明， SPFM V序列与参考序列的同源性92％～97％。

51 samples of sweet potato from Fujian province were detected by the double antibody enzyme-linked immunosorbent method （DAS-ELISA） . The results showed that 24 samples were SPFMV-positive ,the positive rate were 47.06% ;the highest positive rate was 71.40% ,detected in the samples from Quanzhou ,and the next were 50.00% ,detected in samples from Nanan and Longyan .A pair of primer was designed and synthesized according to the published nucleotide sequence of coat protein gene of SPFMV ,and a specific RT-PCR detection procedure was established .The amplified SPFMV-specific fragment in PCR contained 441bps ,and showed 92% -97% ,homology with the SPFMV reference sequence .