检测生物被膜铜绿假单胞菌、克雷伯菌

    产B 2内酰胺酶活性

    李胜岐, 谷 秀, 李乃静, 张智洁, 刘 勇

    (中国医科大学第二附属医院, 辽宁 沈阳 110003)

    摘要: 目的 检测铜绿假单胞菌、克雷伯菌的浮游菌, 生物被膜(biof ilm, BF)菌 B 2内酰胺酶活性; 比较亚胺培南、头孢西丁、哌拉西林诱导这两种BF 菌产酶能力。方法 用改良法建立铜绿假单胞菌、克雷伯菌BF 膜型, 用银染

    法及扫描电镜对BF 进行鉴定; 用紫外分光光度法及B io2 Rad 蛋白定量试剂法检测铜绿假单胞菌、克雷伯菌的浮

    游菌,BF 菌, 亚胺培南、头孢西丁、哌拉西林诱导BF 菌产B 2内酰胺酶的酶活性。结果 铜绿假单胞菌和克雷伯菌

    的两种BF 菌 B 2内酰胺酶活性均显著高于浮游菌(P < 0. 01) , 各种抗生素诱导BF 菌的酶活性显著高于BF 菌(P

    < 0. 01) , 亚胺培南诱导BF 菌产酶活性均显著高于头孢西丁和哌拉西林(P < 0. 01) , 铜绿假单胞菌除浮游菌外各

    组酶活性均显著高于克雷伯菌各组(P < 0. 01)。结论 BF 铜绿假单胞菌、 BF 克雷伯菌能产生大量 B 2内酰胺酶,BF 铜绿假单胞菌产酶能力显著高于BF 克雷伯菌; 亚胺培南诱导BF 菌产酶的能力最强, 其次是头孢西丁, 哌拉

    西林诱导能力最弱。

    关键词: 生物被膜; 铜绿假单胞菌; 克雷伯菌; B 2内酰胺酶; 亚胺培南

    中图分类号: R378. 99+

    1 文献标识码: A 文章编号: 100524529 (2002) 0920649203

    B-Lactama se in Pseudom ona s a erug inosa and Klebs iel la Biof ilm: An Assay

    L I Sheng2 qi, GU Xiu, L IN ai2jing, ZHAN G Zh i2jie, L IU Yong

    (D ep a rtm en t of R esp ira tory D iseases, the S econd C l in ica l Col leg e, Ch ina M ed ica l U n iversity ,S heny ang 110003, Ch ina)

    Abstract : OBJECTIVE To determ ine the act ivity of B 2lactamase of p lank tonic and biof ilm bacter ia of P seu2

    d om onas aerug inosa and K lebsiel la . To compare the ability of im ipenem w ith cefoxit in and p iperacillin that induce

    these two k inds of bacter ia to p roduce B 2lactamase . METHODS In v itro models of P. aerug inosa and K lebsiel la

    bacter ial biof ilm w ere built up in silicon disk w ith the modif ied f lat2 board method and a rap id staining p rocedure

    of A gNO3w as used to ver ify them. The biof ilm w as observed under scanning elect ron m icro scope . The test w as

    developed under th ree group s . Theyw ere p lank tonic bacter ia of P. aerug inosa and K lebsiel la, their biof ilm bacte2

    r ia and biof ilm bacter ia induced by im ipenem, cefoxit in, p iperacillin . The act ivity of B 2lactamase w as quant itated

    by a spect ropho tomet r ic assay method . B 2 L actamase quant itat ion w as determ ined by using the B io2 Rad p ro tein

    assay . RESULTS The act ivity of B 2lactamase of P. aerug inosa and K lebsiel la p lank tonic bacter ia and these two

    k inds of biof ilm bacter ia w ere much h igher than tho se of p lank tonic bacter ia (P < 0. 01). Im ipenem can induce

    h igher B 2lactamase p roduct ion than tho se of cefoxit in and p iperacillin . The act ivity of th ree group s of P. aerug i2

    nosa w as h igher than that of K lebsiel la p rofoundly w ith the stat ist ic method of t2test (P < 0. 01). CONCLU2

    SIONS P. aerug inosa and K lebsiel la biof ilm bacter ia could p roduce a great quant ity of B 2lactamases . The ability of

    P. aerug inosa p roducing B 2lactamase w as st ronger than that of K lebsiel la . Im ipenem w as the st rongest inducer to

    induce biof ilm bacter ia to p roduce B 2lactamase . The second onew as cefoxit in . P iperacillin w as thew eakest induc2

    er .

    Key words : B iof ilm; P seud om onas aerug inosa; K lesiel la; B 2 L actamase; Im ipenem

    生物被膜(BF)菌引起的生物材料相关感染和

    慢性支气管肺感染的难治性1

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