免疫捕捉PCR法检测西瓜细菌性果斑病  

Detection for Pathogen of Bacterial Fruit Blotch of Watermelon by Immuno-capture PCR

作　　者：赵丽涵[1] 王笑[1] 谢关林[1] 徐福寿[2] 谢国雄[2] 

机构地区：[1]浙江大学生物技术研究所,杭州310029 [2]杭州市植保站,杭州310020

出　　处：《农业生物技术学报》2006年第6期946-951,共6页Journal of Agricultural Biotechnology

基　　金：杭州市重大科技项目(No.200432239);国家自然科学基金(No.30370951)资助。

摘　　要：利用免疫吸附富集结合经典PCR技术建立了免疫捕捉PCR检测西瓜(Citrulluslanatus)果斑病菌(Acidovoraxavenaesubsp.citrulli)的检测法,并与直接PCR和生长检测法比较。结果表明,所测果斑病菌产生360bp左右的特异性片段,而10个不同属的对照菌株无特异性片段产生。免疫捕捉PCR检测果斑病菌的灵敏度为50￣102cfu/mL,而直接PCR则为104cfu/mL,两者的灵敏度相差100倍左右。免疫捕捉PCR法对市售7种瓜种的检测发现,其中1种哈密瓜种子、2种甜瓜种子和2种西瓜种子检出果斑病菌,与人工气候箱内生长检测的发病结果基本吻合。显示了该法准确、灵敏、快速、低成本等优点。An immuno-capture PCR （IC-PCR） method for detection of Acidovorax avenae subsp, citrulli, a pathogen of bacterial fruit blotch of watermelon（Citrullus lanatus）, has been established by combination of the fluoride ion-selective electrode（ISE） method with classical PCR technic and compared with the direct PCR method and growth-checking method. The results showed that all the strains of A. avenae subsp, citrulli tested produced 360 bp specific fragments by the IC-PCR and direct PCR method, while other strains of 10 different genera showed negative PCR result. The minimum detection concentration of the IC-PCR and direct PCR method was about 50-10^2 cfu/mL and 10^4 cfu/mL, respectively. The sensitivity of the former was 100 times higher than that of the later. Detection of 7 batches of different melon seeds from the markets by IC-PCR showed that 1 cantaloupe variety, 2 honey melon varieties and 2 watermelon varieties of seeds carried the pathogen, which almost matched with the result of the growth-checking of the melon seeds. It indicates that the IC-PCR is accurate, sensitive, rapid and low cost.

关 键 词：西瓜果斑病菌 检测 PCR 免疫捕捉PCR 

分 类 号：S188[农业科学—农业基础科学]