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选择马铃薯环腐病菌(Clavibacter michiganensis subsp.sepedonicus)基因组DNA特有的保守区域(pCSL0067)设计一套LAMP引物,通过优化反应条件,建立了马铃薯环腐病菌LAMP检测体系。利用多种参比菌的DNA为模板对LAMP检测体系特异性进行了验证,利用马铃薯环腐病菌DNA溶液和菌液梯度稀释液对LAMP检测体系的检测灵敏度进行了验证。在特异性试验中,LAMP检测体系仅对马铃薯环腐病菌进行扩增,对非靶标菌不产生扩增。LAMP检测体系DNA和菌体检测灵敏度分别达到了0.527×10-3 ng/uL和150 CFU/mL。为马铃薯环腐病菌的检测提供了一种新的、简便、快速的检测手段。
Abstract：The specific region of the genome of Clavibacter michiganensis subsp. sepedonicus(pCSL0067) was used for the design of LAMP primers, and the LAMP detection system were established by optimization of reaction conditions. Specificity of the system was verified using a series of reference strains, sensitivity of the system was verified using a series of gradient dilution of DNA solution of the C. michiganensis subsp. sepedonicus and the bacterial suspension. In the specifi city test, LAMP system showed high and identical specifi city. DNA and bacterial suspension sensitivity of LAMP system reached 0.527×10-3 ng/uL and 150 CFU/mL respectively. This study provide a new, fast and easy way for the detection of C.michiganensis subsp. sepedonicus.
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基本信息:
DOI：
中图分类号:S435.32
引用信息:
[1]汪万春,袁钧,郑春生等.马铃薯环腐病菌LAMP检测方法的建立[J].植物检疫,2014,28(01):29-32.
基金信息:
国家“十二五”科技支撑计划(2012BAK11B02)