Haemophilus parasuis is a kind of symbiotic bacteria in upper respiratory tract of pigs,but it can invade the body and cause serious systemic disease under certain conditions.Distinction between virulent strains and avirulent strains is the key point for diagnose of the disease.In order to search for differentially expressed proteins between virulent and attenuated strains of H.parasuis,the proteomics of the cytosolic proteins were compared between H.parasuis virulent 14A7-2 strain and avirulent 20A3-2 strain by twodimensional gel electrophoresis.The cytosolic proteins of two strains were extracted at late logarithmic phase for 2-DE.After electrophoresis,the gels were analyzed with Imagemaster 2D Platinum 7.0 software,and differentially expressed proteins were identified by MALDI-TOF/MS.By 2-DE and analysis,the results showed that there were 586± 10 protein spots in virulent strain and 568± 10 in avirulent strain.A total of 80 protein spots were differentially expressed in the two strains.44 protein spots unique for 14A7-2 or 20A3-2 strain were selected to be identified by mass spectrometry,and 42 protein spots were successfully identified corresponding to 37 proteins.Five proteins were unique for non-virulent strain,while 27 proteins were unique for virulence strains which included ferric uptake regulation protein,trigger factor and 3-dehydroquinate dehydratase associated with virulence.These results provide new information for further studies of virulent factors,pathogenic mechanisms and diagnostic techniques to distinguish different virulence strains ofH.parasuis.%副猪嗜血杆菌(Haemophilus parasuis)是猪群上呼吸道的一种共栖菌群,在一定的条件下能侵入机体并导致严重的全身性疾病,区分出正常栖生菌株和致病性菌株是确诊该病的关键.为了筛选副猪嗜血杆菌毒力株与非毒力株之间的差异蛋白,本研究通过比较蛋白质组学分析临床分离的毒力菌株14A7-2和非毒力菌株20A3-2之间2-DE图谱的差异,提取两个菌株的菌体蛋白进行双向电泳分离,Imagemaster 2D Platinum 7.0软件分析处理凝胶图像,MALDI-TOF/MS鉴定差异蛋白.在副猪嗜血杆菌毒力菌株中检测到596±10个蛋白质点而在非毒力菌株中检测到568±10个蛋白点,其中表达量差异达5倍以上及毒力菌株或非毒力菌株特有的蛋白点共有80个.选取毒力菌株或非毒力菌株特有的44个差异蛋白点进行质谱鉴定,成功鉴定42个蛋白点,对应37种蛋白.其中,毒力菌株特有蛋白27种,非毒力菌株特有蛋白5种,在毒力菌株特有蛋白中鉴定出潜在毒力因子铁摄取调节蛋白、触发因子、3-脱氢奎尼酸脱水酶等.研究结果为进一步研究副猪嗜血杆菌的毒力因子和致病机理以及鉴别诊断不同毒力株新技术提供新信息.
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