铜绿假单胞菌耐药性分析及耐药基因mexA、mexB、oprM的研究

发布时间：2018-11-16 13:24

【摘要】：[目的]分析昆明医科大学第一附属医院铜绿假单胞菌的耐药情况和多重耐药铜绿假单胞菌的感染危险因素,为临床合理选用抗生素及预防多重耐药铜绿假单胞菌感染提供实验室依据;研究铜绿假单胞菌中mexA、mexB及oprM基因的表达,为进一步了解铜绿假单胞菌的耐药机制提供依据。[方法]收集2015年1月1日-2016年12月31日昆明医科大学第一附属医院各科室住院患者送检标本中分离的院内感染铜绿假单胞菌。用VITEK-2全自动药物敏感试验分析仪进行铜绿假单胞菌的药物敏感试验。根据药物敏感试验结果将患者分为多重耐药铜绿假单胞菌感染组(MDRPA)和非多重耐药铜绿假单胞菌感染组(NMDRPA)。回顾性收集铜绿假单胞菌感染患者的临床资料,分析多重耐药铜绿假单胞菌的感染危险因素。逆转录荧光定量聚合酶链反应检测2016年1月1日-2016年12月31日收集的铜绿假单胞菌中编码MexAB-OprM主动外排系统的mexA、mexB及oprM基因。分析mexA、mexB、oprM基因高表达与铜绿假单胞菌对抗生素耐药及多重耐药性之间的关系。[结果]本次研究共收集铜绿假单胞菌242株,其中多重耐药菌株为68株,占28.1%;主要标本来源为呼吸道标本(59.9%),其次是尿液标本(11.2%);242株铜绿假单胞菌主要分布于ICU(24.4%),其次为神经外科(16.1%)。铜绿假单胞菌对抗生素的耐药率由高到低依次为:亚胺培南(36.7%)、美罗培南(30,4%)、头孢他啶(23.8%)、哌拉西林(23.3%)、环丙沙星(20.4%)、妥布霉素(19.9%)、庆大霉素(19.5%)、头孢吡肟(18.8%)、哌拉西林-他唑巴坦(17.8%)、左氧氟沙星(17.6%)、阿米卡星(12.1%)。分离自ICU患者的铜绿假单胞菌耐药率为15.5-57.6%,高于非ICU科室患者分离株(11-29.8%);分离自儿童、成人及老年人患者菌株耐药率分别为0-25%、18.0%-41.4%和4.6%-33.3%。单因素分析表明标本采集前一个月内入住ICU、住院天数、使用氟喹诺酮类抗生素、使用碳青霉烯类抗生素、使用糖肽类抗生素、联合用药、抗生素使用时间、留置尿管、留置胃管、深静脉置管、气管插管、吸痰、使用呼吸机、体外引流、慢性肾功能衰竭及中性粒细胞减少在MDRPA组和NMDRPA组间的分布差异具有统计学意义(P0.05);多因素分析表明碳青霉烯类抗生素和慢性肾功能衰竭在MDRPA组和NMDRPA组间的分布差异具有统计学意义(P0.05)。同时我们的研究显示2016年1月1日-2016年12月31日共收集铜绿假单胞菌102株。102株铜绿假单胞菌中mexA、mexB及oprM基因高表达菌株检出率分别为65.7%、39.2%及35.3%。mexA基因高表达与铜绿假单胞菌对哌拉西林、哌拉西林-他唑巴坦、头孢他啶及头孢吡西的耐药有关;mexA、mexB基因高表达与铜绿假单胞菌的多重耐药性有关。[结论]铜绿假单胞菌对每种抗生素的耐药率各不相同,分离自不同科室、不同年龄段患者的菌株对每种抗生素的耐药率也各不相同,临床医生应根据药物敏感试验结果合理选用抗生素。碳青霉烯类抗生素和慢性肾功能衰竭是多重耐药铜绿假单胞菌感染的独立危险因素。mexA基因的高表达参与铜绿假单胞菌对β-内酰胺类抗生素的耐药,mexA、mexB基因的高表参与铜绿假单胞菌多重耐药性的形成。
[Abstract]:[Objective] To analyze the drug resistance of P. aeruginosa in the First Affiliated Hospital of Kunming Medical University and the risk factors of multiple drug-resistant Pseudomonas aeruginosa, and provide the basis for the rational selection of antibiotics and the prevention of multiple drug-resistant Pseudomonas aeruginosa infection. The expression of mexA, mexB and oprM in P. aeruginosa was studied.[Method] Collect the nosocomial infection of P. aeruginosa in the hospital of the first affiliated hospital of Kunming Medical University on Jan. 1, 2015-December 31, 2016. The drug-sensitive test of P. aeruginosa was performed with the VITEK 2 full-automatic drug-sensitive test analyzer. The patient was divided into multiple drug-resistant Pseudomonas aeruginosa infection groups (MDRPAs) and non-multiple drug-resistant Pseudomonas aeruginosa infection groups (NMDRPAs) according to the drug-sensitive test results. The clinical data of the patients with Pseudomonas aeruginosa were retrospectively collected and the risk factors of the infection of the multiple drug-resistant Pseudomonas aeruginosa were analyzed. The mexA, mexB, and oprM genes of the MexAB-OprM active efflux system were detected in Pseudomonas aeruginosa collected on 1/ 1/ 2016 to 31-Dec-2016 by RT-PCR. The relationship between the high expression of mexA, mexB and oprM and the drug resistance and multiple drug resistance of P. aeruginosa was analyzed.[Results] In this study, 242 strains of P. aeruginosa were collected. The multiple drug-resistant strains were 68 strains, accounting for 28.1%. The main samples were respiratory tract specimens (55.9%), followed by urine specimens (11.2%), 242 strains of P. aeruginosa were mainly distributed in the ICU (2.4%). followed by neurosurgery (16.1%). The resistance rate of Pseudomonas aeruginosa to antibiotics was from high to low in the order of imipenem (32.7%), meropenem (30, 4%), ceftriaxone (23. 8%), methicillin (23. 3%), ciprofloxacin (23.4%), butramycin (19.9%), and gentamicin (19. 5%). Ceftaxime (1.8%), penethamate-other sulbactam (17.8%), levofloxacin (17.6%), and amikacin (12.1%). The drug-resistant rate of P. aeruginosa isolated from ICU patients was 15. 5-55.7%, which was higher than that in non-ICU (11-29. 8%). The drug-resistant rates of the isolates from children, adults and the elderly were 0-25%, 18. 0%-41.4% and 4.6%-33.3%, respectively. The single-factor analysis showed that in the ICU and the number of days of the stay in the ICU, the use of carbapenem antibiotics, the use of the carbapenem antibiotics, the use of glycopeptide antibiotics, combined use, the time of use of antibiotics, the indwelling urinary tube, the indwelling gastric tube, the deep vein tube, the tracheal cannula, The distribution of sputum, ventilator, in-vitro drainage, chronic renal failure and neutropenia was statistically significant between the MDRPA and the NMDRPAs (P0.05). The multi-factor analysis showed that the distribution of carbapenem antibiotics and chronic renal failure in the MDRPA group and the NMDRPA group was statistically significant (P0.05). A total of 102 strains of P. aeruginosa were collected from January 1, 2016 to December 31, 2016. The positive rates of mexA, mexB and oprM genes in 102 strains of P. aeruginosa were 65. 7%, 39. 2% and 35. 3% respectively. The high expression of mexA gene was similar to that of P. aeruginosa. The high expression of mexA and mexB was related to the multiple drug resistance of P. aeruginosa.[Conclusion] The resistance rate of P. aeruginosa to each antibiotic is different, and the drug resistance rate of each antibiotic is different from different departments and different age groups. The clinical doctor should reasonably select antibiotics according to the results of the drug sensitivity test. carbapenem antibiotics and chronic renal failure are independent risk factors for multiple drug-resistant Pseudomonas aeruginosa infection. The high expression of mexA gene was involved in the drug resistance of P. aeruginosa to the antibiotic resistance of P. aeruginosa, and the high expression of mexA and mexB gene was involved in the formation of multiple drug resistance of P. aeruginosa.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R446.5

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