摘要
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.
PCR amplification was performed of the genomic DNA of Xanthomonas campestris pv. begonia (Xcb) and its internal transcribed space (ITS) sequences were obtained. Based on the differences in ITS sequences be- tween Xcb and other Xanthomonas bacteria, one pair species-specific primers XCB(2)/XCB(3) was designed, and the length of the PCR amplified fragment was 400 bp. Thus a new highly efficient means for the detection of X. campestris pv. begonia was developed. In this study, the sensitivity of PCR and serologic test techniques was compared. The PCR detection sensitivity was 27 pg/μL of genomic DNA, which was higher than the serologic test technique.
出处
《西南大学学报(自然科学版)》
CAS
CSCD
北大核心
2014年第7期17-23,共7页
Journal of Southwest University(Natural Science Edition)
基金
农业部公益性行业专项基金资助项目(201303015)
国家自然科学基金资助项目(31360002)
云南省博士新人奖资助项目