替加环素与舒巴坦联合应用对泛耐药鲍曼不动杆菌体外抗菌活性的研究

发布时间：2018-04-05 05:03

本文选题：舒巴坦　切入点：替加环素　出处：《泰山医学院》2013年硕士论文

【摘要】：目的随着医疗技术的发展，大量广谱药物、糖皮质激素、免疫抑制剂的使用，鲍曼不动杆菌已成为院内获得性感染的重要病原菌，尤其是多重耐药、广泛耐药和全耐药鲍曼不动杆菌，由于其快速获得和传播耐药性的能力，近年已呈世界性流行，成为临床治疗的棘手问题。2011年中国CHINET细菌耐药性监测网数据也表明鲍曼不动杆菌感染形势非常严峻，不动杆菌在呼吸道的检出率仅次于大肠埃希菌与肺炎克雷伯菌，鲍曼不动杆菌是引起肺部感染出现率增高较快的细菌，已成为肺部院内感染的主要病原体，它同样也可引发泌尿系感染、继发性脑膜炎、败血症等。鲍曼不动杆菌对重症监护病房（ICU）患者威胁很大，易在ICU引起爆发流行。最近有关英国和美国在伊拉克和阿富汗的军事和非军事人员感染耐药鲍曼不动杆菌的报道进一步说明鲍曼不动杆菌感染的严重性。由于临床感染多为耐药菌株，临床治疗非常困难，甚至是无药可用。目前，对于泛耐药菌株，临床常选择替加环素联合舒巴坦治疗，取得了一定的疗效，但对于两者是否具有协同性尚不清楚，本研究通过观察两者单药及联合的MIC，以其能为临床用药提供科学依据。材料和方法菌株：实验用鲍曼不动杆菌均取自2011年4月至2012年9月济南军区总医院从临床标本中分离并鉴定的广泛耐药耐鲍曼不动杆菌32株（痰标本24株，分泌物4株，引流液2株，脑脊液2株）。标准质控菌株：大肠埃希菌ATCC25922，大肠埃希菌ATCC35218(为监控β内酰胺/β-内酰胺酶抑制剂纸片用)由济南军区总医院细菌室提供。抗菌药物：(1)氨苄西林/舒巴坦、环丙沙星、左氧氟沙星、美洛培南、亚胺培南、庆大霉素、妥布霉素、丁胺卡那、哌拉西林、哌拉西林/他唑巴坦、头孢吡肟、头孢曲松、头孢噻肟、多西环素、米诺环素、复方新诺明、头孢哌酮/舒巴坦的药敏纸片均购自Oxoid赛默飞世尔生物化学制品北京有限公司。（2）进口注射用替加环素，50mg/支辉瑞制药有限公司提供。（3）注射用舒巴坦，0.5g/支齐鲁制药有限公司提供。方法：选取济南军区总医院临床感染标本，常规培养，分离细菌，经VITEK全自动分析仪来鉴定鲍曼不动杆菌，采用纸片扩散法用多种抗生素进行药敏实验，选取广泛耐药鲍曼不动杆菌32株。首先无菌操作将100ul微量肉汤分别加入灭菌的96孔聚苯乙烯板中，然后将替加环素与舒巴坦两种药物分别在96孔板中连续倍比稀释，最后将浓度为3×105CFU/ml的菌悬液100ul加入孔中，孔中菌最终浓度为1.5×105CFU/ml，过夜培养后分别测定替加环素与舒巴坦最小抑菌浓度，再根据两种药物所测得最小抑菌浓度，采用棋盘格微量肉汤稀释法测定（横向为舒巴坦的降梯度方向，则纵向为替加环素的降梯度方向）两药各自的最小抑菌浓度，再根据替加环素与舒巴坦联合用药前后的最小抑菌浓度（MIC）计算部分抑菌浓度（FIC）。最后选取两药表现出协同作用的菌株应用时间杀菌曲线法进一步评价两药的相互作用关系的信息。结果通过WHONET5.4软件中选取经VITEK-2分析仪鉴定32株鲍曼不动杆菌，，再次通过纸片扩散法进行药敏试验，所选取32株不动杆菌均为广泛耐药的鲍曼不动杆菌。17种抗菌药物对32株泛耐药鲍曼不动杆菌的检测结果显示:氨苄西林/舒巴坦、环丙沙星、左氧氟沙星、美洛培南、亚胺培南、庆大霉素、妥布霉素、丁胺卡那、哌拉西林、哌拉西林/他唑巴坦、头孢吡肟、头孢曲松、头孢噻肟、多西环素、米诺环素、复方新诺明、头孢哌酮/舒巴坦耐药率均为100%。舒巴坦对32株XDRAB的MIC范围为32~128ug/ml，MIC90为128ug/ml，替加环素对32株XDRAB的MIC范围为0.5~1ug/ml，MIC90为1ug/ml。舒巴坦和替加环素联用，协同作用28.2%（9/32）、相加作用62.5%（20/32）、无相关作用9.3%（3/32），无拮抗作用。对于棋盘格微量肉汤法所测9株表现为协同的XDR鲍曼不动杆菌，选取舒巴坦、替加环素两药浓度均为0.5×MIC应用时间杀菌曲线法，所选9株XDR鲍曼不动杆菌均表现为协同作用，未见两药联合出现拮抗作用。结论舒巴坦与替加环素联合对XDR鲍曼不动杆菌多表现出协同或相加作用，临床上治疗由XDRAB引起的重症感染，可根据药敏试验采用舒巴坦与替加环素联合应用。
[Abstract]:objective
With the development of medical technology, broad-spectrum drugs, corticosteroids, immunosuppressive drugs, Bauman Acinetobacter has become an important pathogen of nosocomial infection, especially multidrug-resistant, extensively drug-resistant and drug resistance of Acinetobacter Bauman, due to its ability to quickly obtain and spread of drug resistance, in recent years has been in the world pop,.2011 become the thorny issue of clinical treatment of China CHINET bacterial resistance monitoring network data also show that Bauman Acinetobacter infection situation is very serious, Acinetobacter in respiratory rate after Escherichia coli and Klebsiella pneumoniae, Bauman Acinetobacter is caused by lung infection occurrence rate of bacteria has increased faster, become the main pathogens of nosocomial infection in the lungs, it also can cause urinary tract infection, secondary meningitis, septicemia. Bauman Acinetobacter in ICU (ICU) patients The threat of a large, easy to cause the outbreak of ICU. Recently the United Kingdom and the United States in Iraq and Afghanistan military and non military personnel infected with drug-resistant Acinetobacter Bauman reported that Bauman did not move further serious infection. The infection for clinical drug resistant strains, the clinical treatment is very difficult, or even no drugs available. At present, the pan resistant strains, the clinical choice of tigecycline combined with sulbactam treatment has certain curative effect, but both are cooperative is not clear, this study observed both alone and in combination with MIC, which can provide scientific basis for clinical medication. Materials and methods
Strain: the Acinetobacter baumannii from Bauman's laboratory was isolated from the clinical specimens and identified 32 strains of extensively drug-resistant Acinetobacter baumannii from April 2011 to September 2012. There were 24 sputum specimens, 4 secretions, 2 draining fluid and 2 cerebrospinal fluid in the General Hospital of Jinan Military Area, from April 2011 to September 2012.
Standard quality control strain: Escherichia coli ATCC25922, Escherichia coli ATCC35218 (for monitoring beta lactam / beta lactamase inhibitor disk), provided by General Hospital of Jinan Military Area bacteriology room.
Antibiotics: (1) ampicillin / sulbactam, levofloxacin, ciprofloxacin, meropenem, imipenem, gentamicin, tobramycin, amikacin, piperacillin, piperacillin / tazobactam, cefepime, cefotaxime, ceftriaxone, doxycycline, minocycline, compound sulfamethoxazole Ming, Cefoperazone / sulbactam, the drug sensitive slips were purchased from Oxoid thermofisher biochemical products Co. Ltd. of Beijing. (2) imported Tigecycline for Injection, 50mg/ branch of Pfizer Inc to provide. (3) with sulbactam injection, 0.5g/ Qi Lu Pharmaceutical Company Limited.
Methods: specimens of clinical infection, General Hospital of Jinan Military Area, conventional culture, bacterial isolates by VITEK automatic analyzer to identify Bauman Acinetobacter, the drug sensitive experiment was performed with multiple antibiotics disk diffusion method, wide selection of resistant Bauman Acinetobacter 32 strains. First, 96 hole polystyrene board 100ul broth were added to sterilized bacteria then, tigecycline and sulbactam were two drugs in 96 well plate continuous dilution, the concentration of 3 * 105CFU/ml bacterial suspension 100ul into the hole, the hole in the final concentration of bacteria is 1.5 * 105CFU/ml, after overnight incubation of tigecycline and sulbactam minimum inhibition the concentration were determined according to two kinds of drugs measured by minimal inhibitory concentration was determined by the checkerboard microdilution method (transverse gradient direction, sulbactam is longitudinal for tigecycline gradient direction) two drugs The minimum inhibitory concentration respectively, then according to add ring with minimal inhibitory concentration before and after sulbactam combination (MIC) to calculate the fractional inhibitory concentration (FIC). Finally two drugs showed bactericidal curve with strain time method of synergy further evaluation of two drug interactions between information.
Result
Through the WHONET5.4 software in the VITEK-2 analyzer and identification of 32 strains of Bauman Acinetobacter, again drug sensitive test was performed by disk diffusion method, selected 32 strains of Acinetobacter were extensively drug resistant Acinetobacter Bauman.17 antimicrobial agents against 32 strains of Pan resistant Acinetobacter Bauman test results showed: ampicillin / sulbactam sulbactam, levofloxacin, ciprofloxacin, meropenem, imipenem, gentamicin, tobramycin, amikacin, piperacillin, piperacillin / tazobactam, cefepime, cefotaxime, ceftriaxone, doxycycline, minocycline, cotrimoxazole, Cefoperazone / sulbactam are 100%.
Shubatan of 32 strains of XDRAB MIC range of 32~128ug/ml, MIC90 128ug/ml, MIC range of tigecycline against 32 strains of XDRAB 0.5~1ug/ml, MIC90 1ug/ml. Shubatan and tigecycline combined, synergistic effects of 28.2% (9/32), additive effect (20/32), 62.5% no 9.3% (3 /32). No antagonism. The checkerboard microdilution broth method in 9 strains showed a synergistic XDR Bauman Acinetobacter, selected Shubatan, tigecycline two concentration was 0.5 * MIC application time killing curve method, selected 9 strains of Bauman Acinetobacter XDR showed synergistic effect, no two the drug combination of antagonism.
conclusion
Shubatan combined with tegacycline showed more synergistic or synergistic effects on Acinetobacter XDR XDRAB. In clinic, the severe infection caused by XDRAB can be treated in clinic, and the combination of Shubatan and tegacycline should be applied according to the susceptibility test.

【学位授予单位】：泰山医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R563.1

【参考文献】