| 免疫捕捉PCR法检测西瓜细菌性果斑病研究 |
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| 引用本文： | 赵丽涵,王笑,谢关林,徐福寿,谢国雄.免疫捕捉PCR法检测西瓜细菌性果斑病研究[J].农业生物技术学报,2006,14(6):946-951. |
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| 作者姓名： | 赵丽涵 王笑 谢关林 徐福寿 谢国雄 |
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| 作者单位： | 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 |
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| 基金项目： | 杭州市重大科技项目(No.200432239)，国家自然科学基金(No.30370951)资助。 |
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| 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。
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| 关 键 词： | 西瓜果斑病菌 检测 PCR 免疫捕捉PCR |
| 文章编号： | 1006-1304（2006）06-0946-06 |
| 收稿时间： | 2006-07-10 |
| 修稿时间： | 2006-09-06 |
Detection for Pathogen of Bacterial Fruit Blotch of Watermelon by Immuno-capture PCR |
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| ZHAO Li-han,WANG Xiao,XIE Guan-lin,XU Fu-shou,XIE Guo-xiong.Detection for Pathogen of Bacterial Fruit Blotch of Watermelon by Immuno-capture PCR[J].Journal of Agricultural Biotechnology,2006,14(6):946-951. |
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| Authors: | ZHAO Li-han WANG Xiao XIE Guan-lin XU Fu-shou XIE Guo-xiong |
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| Institution: | 1. Institute o f Biotechnology, Zhejiang University, Hangzhou 310029, China; 2. Hangzhou Plant Protection Station, Hangzhou 310020, China |
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| Abstract: | 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. |
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| Keywords: | bacterial fruit blotch of watermelon detection PCR immuno-capture PCR |
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