建兰花叶病毒的分子生物学检测  

Molecular Biology Detection for Cymbidium Mosaic Virus

作　　者：高焕利[1] 杨翠云[2] 于翠[2] 马青[1] 

机构地区：[1]西北农林科技大学植保学院,陕西省农业分子生物学重点实验室,陕西杨凌712100 [2]上海出入境检验检疫局,上海200135

出　　处：《西北农业学报》2008年第2期289-292,296,共5页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：上海市科委技术标准专项项目(04DZ05064);教育部长江学者和创新团队发展计划项目(No.200558)

摘　　要：根据建兰花叶病毒外壳蛋白基因的保守序列设计2对引物,采用IC-RT-PCR方法和质粒梯度稀释的方法,研究建兰花叶病毒的分子检测方法和灵敏度。结果表明,从21个建兰花叶病毒的病株样品中均能扩增出与预期大小相同的DNA条带,序列测定和序列同源性分析表明所测定的序列均为CyMV外壳蛋白基因的部分序列,序列同源性均达到89%以上;灵敏度检测结果表明:可检测的最低病毒含量为2.72×10-7μg/mL。Two pairs of primers specific to the conserved sequence of Cymbidium mosaic virus （CyMV） coat protein gene were designed, the IC-RT-PCR and different dilution of plasmind of CyMV were used to study the molecular detection method and the sensitivity for CyMV. A specific DNA band of expected size was amplificated from all the 21 isolates of CyMV. Sequencing analysis showed that the determined sequence was partial CyMV coat protein gene and their homology reached 89%. The sensitivity of CyMV was determined using dilution of plasmind of CyMV and the minimum content of virus inspected was 2.72×10.7μg/mL.

关 键 词：建兰花叶病毒(CyMV) 免疫捕获反转录聚合酶链式反应 灵敏度检测 

分 类 号：S436.8[农业科学—农业昆虫与害虫防治]