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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。
A LAMP primer was designed based on the conserved region (pCSL0067) of genomic DNA of Clavibacter michiganensis subsp. Sepedonicus. By optimizing the reaction conditions, a LAMP detection system was established for ring rot fungi. The specificity of LAMP detection system was verified by using a variety of reference bacteria DNA as a template. The detection sensitivity of LAMP detection system was verified by using DNA solution of bacterial ring rot and bacterial gradient dilution. In the specificity test, the LAMP detection system only amplifies the P. annua and does not amplify the non-target bacteria. LAMP detection system DNA and bacterial detection sensitivity reached 0.527 × 10-3 ng / uL and 150 CFU / mL. It provides a new, simple and rapid detection method for the detection of ringworm in potato.