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对丽格海棠细菌性叶斑病菌基因组DNA进行PCR扩增,获得其ITS序列.依据该病菌与该属其他细菌ITS序列的差异,设计特异性引物对XCB(2)/XCB(3),其扩增片段为400bp,由此建立了快速高效检测该病菌的PCR技术体系.研究根据抗原抗体特异性结合的特点,制备了兔疫细菌抗体血清,建立了体外检测病原细菌的血清学方法如琼脂糖双向扩散法及间接ELISA法,为丽格海棠细菌性叶斑病的诊断提供了新的重要手段.该研究还比较了PCR和血清学检测技术的灵敏度差异,结果显示PCR技术检测灵敏度可达27pg/μL,高于血清学检测技术.
According to the difference of ITS sequences between the bacteria and other bacteria in the genus, the specific primer pair XCB (2) / XCB (3) was designed and its genomic DNA was amplified by PCR from genomic DNA of Bacillus lichenotori The amplified fragment was 400bp, thus establishing a rapid and efficient detection of the bacteria PCR technology system.According to the characteristics of the antigen-specific binding, the preparation of the rabbit immune antibody serum, the establishment of in vitro detection of pathogenic bacteria serological methods such as agar The two-dimensional sugar diffusion method and the indirect ELISA method provided a new important method for the diagnosis of Bacterial leaf spot in Begonia species. The sensitivity of PCR and serological detection techniques was also compared. The results showed that the detection sensitivity of PCR was up to 27pg / μL, higher than serological detection techniques.