摘要： 目的 分析铜绿假单胞菌临床分离株生物膜形成能力,探讨生物膜与感染反复发作之间的关系。 方法 收集痰液标本,常规方法分离鉴定得到28株铜绿假单胞菌;应用微孔法测定28株铜绿假单胞菌生物膜形成情况;应用硅胶薄片法建立体外生物膜模型,1%复红染色后观察不同生长时期的生物膜形态变化情况,同时应用平板培养法测定生物膜内活菌数目。 结果 28株铜绿假单胞菌临床分离株中,22株形成生物膜,生物膜阳性率为78.57%(22/28);铜绿假单胞菌临床分离株体外生物膜成熟过程分为4个阶段:第1d为黏附阶段,第2 d为过渡阶段,第3~6d为成熟阶段,第6~10 d为解体阶段;在生物膜成熟过程中,生物膜内活菌数量可发生周期性改变。 结论 铜绿假单胞菌生物膜形成是下呼吸道感染反复发作的重要因素之一。

Abstract: Objective To inspect the capability of biofilm formation by pseudomonas aeruginosa isolated from lower respiratory tract infections,and examine the relationship between biofilm of pseudomonas aeruginosa and recurrent infection. Method After 28 strains of pseudomonas aeruginosa were isolated from sputum samples by routine method.The capability of biofilm formation was examined by micro-cell plate.The biofilm model was established on small silica gel slice in vitro, and the morphology changes of biofilm were observed by light microscope after 1% azaleine staining.Meanwhile,the number of living bacteria in biofilm was calculated by plate culture method. Result In 28 strains of pseudomonas aeruginosa,22 strains produced biofilm,and the positive rate of biofilm was 78157% (22/28).The course of biofilm development in vitro was that,on first day,the bacteria attached the surface of silica gel slice and partly formed small cluster;on second day,the small cluster expanded;on third day and sixth day,the mature membrane-like structure appeared;on tenth day,with membrane-like structure partly destruction,small cluster appeared again.The number of living bacteria in biofilm changed as follows,slowly decreased during first day to second day,then sharply falled during second day to third day,retained a stabilization level during second day to third day,and slowly increased again during sixth day to tenth day. Conclusion In vitro,the biofilm development of pseudomonas aeruginosa contains four stages,attachment phase during first day,transition phase during second day,maturity phase during third day to sixth day,and destruction phase during sixth day to tenth day.In the process of biofilm development,the number of living bacteria in biofilm changes with four stages,and can be used to evaluate the phase of biofilm.The biofilm formation of pseudomonas aeruginosa is an important factor of recurrent infection.