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
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摘要: 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SPFMV呈阳性, 阳性率为47.06%, 其中泉州样品SPFMV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SPFMV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SPFMV的检测方法。该检测方法能够扩增出SPFMV特异性片段, 片段大小为441bp, 测序结果表明, SPFMV序列与参考序列的同源性92%97%。Abstract: 51samples of sweet potato from Fujian province were detected by the double antibody enzyme-linked immunosorbent method (DAS-ELISA) .The results showed that 24samples 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.