中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2010年
11期
1048-1052
,共5页
李小燕%卓超%金光耀%黎晓强%肖书念%钟南山
李小燕%卓超%金光耀%黎曉彊%肖書唸%鐘南山
리소연%탁초%금광요%려효강%초서념%종남산
铜绿假单胞菌%OprD2蛋白%碳青霉烯类药物耐药
銅綠假單胞菌%OprD2蛋白%碳青黴烯類藥物耐藥
동록가단포균%OprD2단백%탄청매희류약물내약
Pseudomonas aeruginosa%OprD porin protein%Carbapenem resistance
目的 研究铜绿假单胞菌外膜通道蛋白OprD2的表达减弱或缺失,以及OprD2蛋白自身突变是否会影响铜绿假单胞菌对碳青霉烯类药物的耐药性.方法 收集分离自临床对亚胺培南(IPM)的最低抑菌浓度(MIC)值≥8μg/m1的铜绿假单胞菌共101株,采用肉汤稀释法检测菌株对比阿培南(BPM)、美罗培南(MEM)、帕尼培南(PEM)的MIC值;荧光定量RT-PCR检测铜绿假单胞菌膜通道蛋白oprD2基因的表达量情况;针对oprD2相对表达量正常并对亚胺培南耐药的铜绿假单胞菌,采用普通PCR的方法扩增oprD2全长基因并测序.结果 根据铜绿假单胞菌的OprD2蛋白相对表达量结果,将101株铜绿假单胞菌分成两组,组1为OprD2相对表达量降低组;组2为OprD2相对表达量正常组;组1与组2相比,对IPM、BPM、MEM、PEM的MIC值≥16μg/ml的菌株比例差异均有统计学意义(P＜0.01).组1中,28株同时对4种药物的MIC均≥16 μg/ml,其中有25株的OprD2的相对表达量明显减低(＜0.4);外膜孔蛋白OprD2转录水平与4种碳青霉烯类药物MICs之间呈负相关趋势.组2中,有16株9prD2基因发生突变,按照突变的类型主要分成4组;与PAO1相比,这些菌株对IPM、BPM、MEM、PEM的MIC值有不同程度的增加.结论 OprD2外膜蛋白的表达量减少或缺失是铜绿假单胞菌对亚胺培南耐药的主要机制,可能也与其他3种碳青霉烯类药物耐药有密切关系;铜绿假单胞菌的oprD2基因发生突变,可能会降低铜绿假单胞菌对这儿种碳青霉烯类药物的敏感性.
目的 研究銅綠假單胞菌外膜通道蛋白OprD2的錶達減弱或缺失,以及OprD2蛋白自身突變是否會影響銅綠假單胞菌對碳青黴烯類藥物的耐藥性.方法 收集分離自臨床對亞胺培南(IPM)的最低抑菌濃度(MIC)值≥8μg/m1的銅綠假單胞菌共101株,採用肉湯稀釋法檢測菌株對比阿培南(BPM)、美囉培南(MEM)、帕尼培南(PEM)的MIC值;熒光定量RT-PCR檢測銅綠假單胞菌膜通道蛋白oprD2基因的錶達量情況;針對oprD2相對錶達量正常併對亞胺培南耐藥的銅綠假單胞菌,採用普通PCR的方法擴增oprD2全長基因併測序.結果 根據銅綠假單胞菌的OprD2蛋白相對錶達量結果,將101株銅綠假單胞菌分成兩組,組1為OprD2相對錶達量降低組;組2為OprD2相對錶達量正常組;組1與組2相比,對IPM、BPM、MEM、PEM的MIC值≥16μg/ml的菌株比例差異均有統計學意義(P＜0.01).組1中,28株同時對4種藥物的MIC均≥16 μg/ml,其中有25株的OprD2的相對錶達量明顯減低(＜0.4);外膜孔蛋白OprD2轉錄水平與4種碳青黴烯類藥物MICs之間呈負相關趨勢.組2中,有16株9prD2基因髮生突變,按照突變的類型主要分成4組;與PAO1相比,這些菌株對IPM、BPM、MEM、PEM的MIC值有不同程度的增加.結論 OprD2外膜蛋白的錶達量減少或缺失是銅綠假單胞菌對亞胺培南耐藥的主要機製,可能也與其他3種碳青黴烯類藥物耐藥有密切關繫;銅綠假單胞菌的oprD2基因髮生突變,可能會降低銅綠假單胞菌對這兒種碳青黴烯類藥物的敏感性.
목적 연구동록가단포균외막통도단백OprD2적표체감약혹결실,이급OprD2단백자신돌변시부회영향동록가단포균대탄청매희류약물적내약성.방법 수집분리자림상대아알배남(IPM)적최저억균농도(MIC)치≥8μg/m1적동록가단포균공101주,채용육탕희석법검측균주대비아배남(BPM)、미라배남(MEM)、파니배남(PEM)적MIC치;형광정량RT-PCR검측동록가단포균막통도단백oprD2기인적표체량정황;침대oprD2상대표체량정상병대아알배남내약적동록가단포균,채용보통PCR적방법확증oprD2전장기인병측서.결과 근거동록가단포균적OprD2단백상대표체량결과,장101주동록가단포균분성량조,조1위OprD2상대표체량강저조;조2위OprD2상대표체량정상조;조1여조2상비,대IPM、BPM、MEM、PEM적MIC치≥16μg/ml적균주비례차이균유통계학의의(P＜0.01).조1중,28주동시대4충약물적MIC균≥16 μg/ml,기중유25주적OprD2적상대표체량명현감저(＜0.4);외막공단백OprD2전록수평여4충탄청매희류약물MICs지간정부상관추세.조2중,유16주9prD2기인발생돌변,안조돌변적류형주요분성4조;여PAO1상비,저사균주대IPM、BPM、MEM、PEM적MIC치유불동정도적증가.결론 OprD2외막단백적표체량감소혹결실시동록가단포균대아알배남내약적주요궤제,가능야여기타3충탄청매희류약물내약유밀절관계;동록가단포균적oprD2기인발생돌변,가능회강저동록가단포균대저인충탄청매희류약물적민감성.
Objective To investigate the impact on the resistance of carbapenem with the expression of OprD2 or OprD2 mutation in Pseudomonas aeruginosa. Methods One hundred and one clinical strains of Pseudomonas aeruginosa with MIC for imipenem ≥8 μg/ml were studied. MIC were determined by the broth microdilution method, and the antibiotics tested were imipenem(IPM ), biapenem( BPM), meropenem(MEM) and panipenem(PEM). The expression of the oprD2 gene in Pseudomonas aeruginosa were analyzed by real-time reverse transcriptase PCR(RT-PCR). For the Pseudomonas aeruginosa with normal expression of OprD2 and resistance to imipenem, full-length oprD2 gene was amplified by PCR and the products were sequenced. Results According to the result of the expression of oprD2 gene, 101 strains of Pseudomonas aeruginosa were divided into two groups: group1 with diminished expression of OprD2, and group2 with normal expression of OprD2. Comparing isolates with MIC of 4 kinds of carbepenem agents ≥ 16 μg/ml in two groups. Data showed the amount of OprD2 expression were different between two groups(P ＜0.01 or P ＜ 0.05). In group1, there are 28 isolates with MIC ≥ 16 μg/ml of all the 4 kinds of carbapenems, among which 25 isolates have obviously diminished expression of OprD2 ( ＜ 0.4). Negative correlations tendency appeared between the level of OprD2 transcription and MICs of 4 kinds of carbepenem agents in Pseudomonas aeruginosa. In group2, 16 strains with OprD2 mutation divided into 4 types according to the pattern of alteration. Compared with PAO1, these strains have increased MIC with different degree to IPM,BPM, MEM and PEM. Conclusion The deletion or diminished expression of OprD2 resulted in resistance to imipenem in Pseudomonas aeruginosa. The level of OprD2 transcription and antimicrobial activities for carbapenem agents proved to be highly correlated in Pseudomonas aeruginosa. The mutation of OprD2 in Pseudomonas aeruginosa probably decreased the sensitivity of carbapenem agents against Pseudomonas aeruginosa.
