布鲁氏菌强毒株与弱毒株基因组DNA差异基因筛选与鉴定

Screening and identification of the differential genomic DNA between Brucella abortus virulent strain and attenuated strain

本研究应用代表性差异分析技术(RDA)对流产布鲁氏菌强毒株544A和弱毒株104M进行基因组差减分析。经过3轮差减杂交,获得强毒株特有的9条差异片段和弱毒株特异的17条差异片段。经BLAST分析和PCR鉴定,最终证实3条差异片段,与流产布鲁氏菌经典弱毒株S19同源性在98%～99%之间,只在弱毒株104M基因组中检测出,而在强毒株544A中不存在。该实验为建立布鲁氏菌自然感染菌与疫苗株的基因鉴别诊断方法奠定了基础。

The differential genomic DNA between Brucella abortus virulent strain 544 A and Brucella abortus attenuated strain 104 M was screened using representational difference analysis （RDA）. Nine differential fragments in virulent strain 544 A and 17 differential fragments in attenuated strain 104 M were identified after three rounds of subtractive hybridization. Three differential fragments detected only in the attenuated strain 104 M, but not in virulent strain 544 A shared 98 % to 99 % homology with the classic Brucella abortus attenuated strain S19 in BLAST. It is possible to use these 3 differential fragments to establish a differential diagnosis for discriminating the Brucella infection from vaccination.