| 利用免疫磁性分离–PCR检测安祖花细菌性枯萎病病原菌 |
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| 引用本文： | 王钊,储丽红,赵凯,彭佳佳,明军,袁素霞,刘春.利用免疫磁性分离–PCR检测安祖花细菌性枯萎病病原菌[J].园艺学报,2013,40(8):1600-1599. |
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| 作者姓名： | 王钊 储丽红 赵凯 彭佳佳 明军 袁素霞 刘春 |
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| 作者单位： | （1中国农业科学院蔬菜花卉研究所，北京 100081；2南京林业大学风景园林学院，南京 210037；3宁夏大学农学院，银川 750021） |
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| 基金项目： | 北京市科技计划研发攻关项目，中国农业科学院与北京市大兴区科技合作项目，农业部园艺作物生物学与种质创制重点实验室项目 |
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| 摘 要： | 根据地毯草黄单胞菌花叶万年青致病变种（Xanthomonas axonopodis pv. dieffenbachiae）的RAPD序列设计特异性引物，并对羧基化磁珠的结合性能进行检验，从而建立和优化了免疫磁性分离–PCR体系，对安祖花细菌性枯萎病进行早期检测。结果表明：1 mg磁珠对多克隆抗体最大吸附值为0.268 mg，进行免疫磁性捕获时免疫磁珠的最佳浓度是0.566 ~ 0.741 mg ?mL-1。免疫磁性分离–PCR可以减少PCR反应中的抑制物质，对X. axonopodis pv. dieffenbachiae的检测灵敏度可以达到10 ~ 100 cfu ?mL-1，比常规PCR检测灵敏至少100倍。
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| 关 键 词： | 安祖花 地毯草黄单胞菌花叶万年青致病变种 免疫磁性分离–PCR 最大吸附值 捕获率 |
| 收稿时间： | 2013-03-18 |
Detection of Xanthomonas axonopodis pv.dieffenbachiae in Authurium andreanum by Immunomagnetic Separation-PCR |
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| WANG Zhao,CHU Li-hong,ZHAO Kai,PENG Jia-jia,MING Jun,YUAN Su-xia,LIU Chun,.Detection of Xanthomonas axonopodis pv.dieffenbachiae in Authurium andreanum by Immunomagnetic Separation-PCR[J].Acta Horticulturae Sinica,2013,40(8):1600-1599. |
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| Authors: | WANG Zhao CHU Li-hong ZHAO Kai PENG Jia-jia MING Jun YUAN Su-xia LIU Chun |
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| Institution: | （1Institute of Vegetables and Flowers，Chinese Academy of Agricultural Sciences，Beijing 100081，China；2College of Landscape Architecture，Nanjing Forestry University，Nanjing 210037，China；3College of Agriculture，Ningxia University，Yinchuan 750021，China） |
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| Abstract: | According to the RAPD of Xanthomonas axonopodis pv. dieffenbachiae，specific primers were designed，and binding ability of carboxyl magnetic beads were tested. The system of optimization of immunomagnetic separation-PCR（IMS-PCR）protocol for detecting the Bacterial Blight of Anthurium were establisted. The results showed that the saturate absorption of 1 mg carboxyl magnetic beads to the polyclonal antibodys is 0.268 mg. The optimal concentration of immunomagnetic beads（IMB）is 0.566–0.741 mg ?mL-1 when X. axonopodis pv. dieffenbachiae is captured. The detection sensitivity for X. axonopodis，by IMS-PCR which can reduce the inhibitory substances in reaction，is 10–100 cfu ? mL-1. It is 100 times more sensitive than ordinary PCR at least. pv. dieffenbachiae |
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| Keywords: | Authurium andreanum Xanthomonas axonopodis pv dieffenbachiae IMS-PCR sensitivity saturation absorption capture rate |
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