免疫捕捉PCR法检测西瓜细菌性果斑病研究

利用免疫吸附富集结合经典PCR技术建立了免疫捕捉PCR检测西瓜果斑病菌的检测法，并与直接PCR和生长检测法作了比较。结果表明，所测果斑病菌Acidovorax avenae subsp. citrulli都产生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则在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。

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

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 50~102 cfu/mL and 104 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.