环二鸟苷酸对铜绿假单胞菌的生物膜形成及消毒剂抗性的影响

Effect of cyclic diguanosine monophosphate on biofilm formation and disinfectant resistance of Pseudomonas aeruginosa

目的 探究C-di-GMP对临床内镜野生型铜绿假单胞菌(PA)生物膜形成能力的影响和对高水平消毒剂的抗性。方法 收集江西省某三甲医院临床内镜上分离的PA,经C-di-GMP诱导后,观察PA在不同时间的生长情况;结晶紫染色法评价C-di-GMP诱导后PA的生物膜形成能力;最小抑菌浓度和最小杀菌浓度试验检测C-di-GMP诱导6 d后的PA生物膜对2.4%戊二醛的抗性。结果 C-di-GMP诱导对野生型PA生长速度没有显著影响(P>0.05),但对生物膜形成能力具有促进作用(P<0.05),且浓度在20μmol/L时生物膜形成能力最强。C-di-GMP诱导6 d后形成的生物膜对消毒剂抗性增强,且浓度在20μmol/L时最小抑菌浓度试验和最小杀菌浓度试验结果高达14 000、24 000 mg/L,消毒剂抗性程度最高。结论 C-di-GMP对临床内镜收集的野生型PA的生物膜形成具有促进作用,并且存在一定量效关系,能增强生物膜对消毒剂的抗性。

Objective To explore the influence of C-di-GMP on the biofilm formation ability and high level of disinfectant resistance of clinical endoscopic wild-type pseudomonas aeruginosa(PA).Methods PA isolated from clinical endoscopy in a grade A hospital in Jiangxi province was collected, and the growth of PA was observed by adding C-di-GMP. The biofilm formation ability of PA induced by C-di-GMP was evaluated by crystal violet staining. The resistance degree of PA biofilm to 2.4% glutaraldehyde was detected by the minimum inhibitory and the minimum bactericidal concentration test after induced by different concentrations of C-di-GMP for 6 days.Results There was no significant effect on the growth rate of wild-type PA(P>0.05), but the biofilm formation ability was promoted with statistical significance(P<0.05), and the biofilm formation ability was strongest at the concentration of 20 μmol/L. The resistance of biofilm to disinfectants was enhanced after 6 days induced by C-di-GMP, and the results of minimum inhibitory concentration test and minimum bactericidal concentration test were as high as 14 000 and 24 000 mg/L when the concentration was 20 μmol/L, indicating the highest degree of disinfectant resistance.Conclusion C-di-GMP can promote the formation of biofilm of wild-type PA collected by clinical endoscopy with a certain effect-dose relationship, and can also enhance the resistance of biofilm to disinfectant.