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目的探讨黏液型与非黏液型铜绿假单胞菌(Pseudomonas aeruginosa,P.a)的差异蛋白谱,以发现黏液型P.a潜在的生物标志物。方法采用来自于临床囊性纤维化患者分离的3株黏液型P.a菌株,脉冲场凝胶电泳以确定它们的同源性;采用同位素相对和绝对定量标记(isobaric tags for relative and absolute quantitation,i TRAQ)结合二维液相色谱串联质谱(two-dimensional liquid chromatography-tandem mass spectrometry,2D LC-MS/MS)技术分析3株黏液型与对照组1株非黏液型P.a的差异蛋白谱,并采用RT-q PCR方法对5个共同差异蛋白进行验证。结果 3株黏液型P.a菌株不具有同源性,i TRAQ结果没有明显差异。3组共有219个上调和101个下调差异蛋白。pa2815、pa2018、dnak、pa5046和acef基因转录水平与i TRAQ结果一致。结论不同黏液表型P.a的i TRAQ结果没有明显差异,219个上调和101个下调差异蛋白可以作为研究黏液型与非黏液型P.a致病机制的候选生物标志物。
Objective To investigate the differential protein profiles of mucoid and non-mucoid Pseudomonas aeruginosa (P.a), and find out the potential biomarkers of mucinous P.a. Methods Three strains of Myxoid Pa isolated from patients with clinical cystic fibrosis were subjected to pulsed-field gel electrophoresis to determine their homology. Isobaric tags for relative and absolute quantitation (i TRAQ ) Was used to analyze the differential protein profiles of three non-mucinous Pa strains in three mucus-type and two control groups by two-dimensional liquid chromatography-tandem mass spectrometry (2D LC-MS / MS) -q PCR method to verify five common differential proteins. Results Three strains of myxomatous P.a did not show homology, and the results of i TRAQ showed no significant difference. There were 219 up-regulated and 101 down-regulated proteins in 3 groups. The pa2815, pa2018, dnak, pa5046 and acef gene transcription levels were consistent with i TRAQ results. Conclusions There is no significant difference in i TRAQ between different mucoid phenotypes. 219 up - regulated and 101 down - regulated proteins can be used as candidate biomarkers to study the pathogenesis of mucinous and non - mucoid.