利用免疫磁性分离–PCR检测安祖花细菌性枯萎病病原菌

园艺学报 ›› 2013, Vol. 40 ›› Issue (8): 1600-1599.

利用免疫磁性分离–PCR检测安祖花细菌性枯萎病病原菌

王钊1，储丽红2，赵凯1，彭佳佳3，明军1，袁素霞1，刘春1,*

（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）

收稿日期:2013-03-18 出版日期:2013-08-25 发布日期:2013-08-25

Detection of Xanthomonas axonopodis pv. dieffenbachiae in Authurium andreanum by Immunomagnetic Separation-PCR

WANG Zhao1，CHU Li-hong2，ZHAO Kai1，PENG Jia-jia3，MING Jun1，YUAN Su-xia1，and LIU Chun1,*

（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）

Received:2013-03-18 Online:2013-08-25 Published:2013-08-25

摘要/Abstract

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。

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

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

Key words: Authurium andreanum, Xanthomonas axonopodis pv. dieffenbachiae, IMS-PCR, sensitivity, saturation absorption, capture rate

王钊1，储丽红2，赵凯1，彭佳佳3，明军1，袁素霞1，刘春1,*. 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.

WANG Zhao1，CHU Li-hong2，ZHAO Kai1，PENG Jia-jia3，MING Jun1，YUAN Su-xia1，and LIU Chun1,*. 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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