耶尔森菌不同成膜基因对其生物膜形成的影响

Effect of different biofilm-forming genes on Yersinia biofilm

【目的】研究耶尔森菌生物膜的检测方法和不同成膜基因,为探讨耶尔森菌不同成膜基因对其生物膜形成的影响提供参考。【方法】应用静置试管悬浮菌膜观察、微孔96孔聚苯乙烯板结晶紫染色法、刚果红平板培养3种方法,检测了25株耶尔森菌生物膜的形成情况;用PCR技术检测了生物膜形成相关基因metE、ompR、rpoS、rhaH在不同菌株中的分布情况,并分析了生物膜形成与这些基因的相关性。【结果】静置试管悬浮菌膜观察、微孔96孔聚苯乙烯板结晶紫染色法和刚果红平板培养法3种方法的耶尔森菌生物膜阳性检出率分别为60%,56%和68%,3种方法检测均为阳性的菌株有11株,均为阴性的有5株,一致率为64%;4种成膜基因在25株耶尔森菌中的检出率分别为52%,56%,32%和24%。利用SPSS分析了4种基因对生物膜形成的影响,其中metE、ompR基因是生物膜形成的主效应因子,二者的累计方差贡献率达到75.149%。【结论】刚果红平板培养法检测生物膜最敏感;metE、ompR、rpoS、rhaH4种基因对生物膜的形成均有不同程度的影响,其中metE、ompR与生物膜的形成密切相关。

[Objective] The research was to study the detection methods for Yersinia biofilm and differ- ent biofilm-forming genes, and the role of different biofilm-forming genes in Yersinia biofilm formation. [Method] Three different methods, including observation of suspended biofilm on the glass test tubes, crystal violet stain in polystyrene microtiter plate and clone observation in Congo red medium, were used to detect biofilms of 25 Yersinia strains, then the distribution of four biofilm-forming genes （metE, ompR, rpoS and rhaH） were detected by PCR method. The correlation between four biofilm-forming genes and Yersinia strains biofilm-forming was analyzed as well. [Result] The results of three methods had a con- cordance of 64%o and 11 Yersinia strains were verified positive and 5 negative among three assays. The dis- tribution of four biofilm-forming genes （metE,ompR,rpoS and rhaH） were 52% ,56% ,32% ,and 24% ,re- spectively. SPSS software found that mete and ompR genes were the main factors with a variance contribu- tion rate of 75%/00. [Conclusion] Congo red medium was the most sensitive method with a positive rate of 68 %. metE,ompR, rpoS and rhaH genes had different effects on the formation of biofilm, especially mete and ompR genes related closely to the biofilm-forming.