舒巴坦联合9种抗菌药物对耐碳青霉烯类鲍曼不动杆菌体外抗菌活性研究

目的：评价舒巴坦与9种临床常用抗菌药物联合用药对耐碳青霉烯类鲍曼不动杆菌（CRAB）的体外抗菌效应。方法：从临床分离得到24株CRAB。采取微量肉汤稀释法测定10种抗菌药物单一用药对CRAB的最低抑菌浓度（MIC）。采用棋盘法设计,微量肉汤稀释法测定舒巴坦分别联合9种抗菌药物对CRAB的MIC,并计算部分抑菌浓度指数（FIC）判定联合效应。结果：9种抗菌药物联用舒巴坦后的MIC均比单用时降低;联用后替加环素、多西环素、环丙沙星、左氧氟沙星的MIC₅₀均低于对应单药敏感折点,替加环素的MIC₉₀达到单药的敏感折点。舒巴坦与9组联合用药均未出现拮抗作用;舒巴坦联合阿米卡星的协同作用最强,其次为美罗培南、帕尼培南、亚胺培南、多西环素、替加环素。结论：与单药相比,9种药物联用舒巴坦对CRAB的体外抑菌作用均有增强效果。9种抗菌药物联合舒巴坦有效地降低MIC值,其中多西环素、替加环素、环丙沙星和左氧氟沙星,可使其MIC值降至敏感范围内。     

EDTA-2Na联合抗菌药物对铜绿假单胞菌的体外抑菌及生物膜抑制作用研究

目的 探讨乙二胺四乙酸二钠(ethylenediaminetetraacetic acid disodium,EDTA-2Na)联合抗菌药物对铜绿假单胞菌的体外抑菌及生物膜抑制作用。方法 对临床分离的10株亚胺培南耐药铜绿假单胞菌、8株黏菌素耐药铜绿假单胞菌和8株阿米卡星耐药铜绿假单胞菌,通过微量肉汤稀释棋盘法检测EDTA-2Na分别与亚胺培南、黏菌素和阿米卡星联用以及各自单用时对相应耐药菌株的最小抑菌浓度(minimal inhibititory concentration,MIC),并通过计算部分抑菌浓度指数(fractional inhibitory concentration index,FICI)评价联合抑菌效果;结晶紫生物膜试验检测EDTA-2Na分别与亚胺培南、黏菌素和阿米卡星联用及各自单用时对相应耐药菌株生物膜的抑制作用,通过测量A595值评价抑制生物膜效果。结果 EDTA-2Na与亚胺培南、黏菌素和阿米卡星联用后对相应耐药菌株的抑菌作用表现为协同或相加,对生物膜也具有明显的协同抑制作用。结论 EDTA-2Na联合抗菌药物对铜绿假单胞菌具有良好的体外抑菌作用及抑制生物膜作用。     

替加环素、环丙沙星、阿米卡星三联对耐碳青霉烯类鲍曼不动杆菌的抑菌效果观察

目的：探讨替加环素（TGC）、环丙沙星（CIP）、阿米卡星（AMK）三联抑制耐碳青霉烯类鲍曼不动杆菌（CRAB）的效果。方法：选取2017年1月－12月河南省新乡市中心医院收集的80株CRAB菌株用于实验，棋盘稀释法测定替加环素、环丙沙星、阿米卡星联合对CRAB的最低抑菌浓度（MIC），计算部分抑菌浓度指数（FICI）和分级生物膜清除浓度指数（FECI）。结果：（1）TGC单用时MIC₅₀、MIC₉₀均高于联用时，敏感率均低于联用时；AMK单用时MIC₅₀、MIC₉₀均高于联用时，敏感率均低于联用时；CIP单用时，MIC₅₀均高于联用时，MIC₉₀与TGC、CIP联用相同，但高于三联用药时；CIP单用时敏感率与TGC、CIP联用相同，但低于三联用药时；（2）协同抑菌率较高的是TGC+AMK+CIP三联，其次是TGC+AMK，TGC+CIP表现为不相关；全部联合组合未发现拮抗现象；（3）TGC+AMK、TGC+CIP、TGC+AMK+CIP方案FECI≤0.5比例分别为7.50%、31.25%、32.50%，三联方案最高；TGC+AMK、TGC+CIP、TGC+AMK+CIP均具有生物膜清除协同效应，TGC+CIP、TGC+AMK+CIP未发现拮抗现象。结论：TGC、AMK、CIP三联较两联、单独用药对CRAB抑菌效果及生物膜清除均更有优势。     

替加环素联合5种抗菌药物对多重耐药鲍曼不动杆菌的体外活性

目的 评价替加环素分别联合5种临床常用抗不动杆菌属抗菌药物对57株多重耐药鲍曼不动杆菌的体外抗菌作用.方法 琼脂棋盘稀释法测定替加环素分别联合美罗培南、阿米卡星、环丙沙星、粘菌素、舒巴坦对57株对美罗培南、阿米卡星、环丙沙星、米诺环素均耐药的多重耐药鲍曼不动杆菌的最低抑菌浓度(MIC),并计算部分抑菌浓度指数(FICI).结果 替加环素与5种抗菌药物联合后表现为协同或不相关作用,其中协同率较高的组合为替加环素+阿米卡星组,50.9％;其次为替加环素+美罗培南组,29.8％,未发现拮抗现象.结论 替加环素与5种抗菌药物联合对本组多重耐药鲍曼不动杆菌主要表现为不相关作用,但与阿米卡星联合具有相对较高的协同率.     

替加环素和多黏菌素对产KPC酶肺炎克雷伯菌的抗菌效应研究

目的 评价替加环素、多黏菌素对产KPC酶肺炎克雷伯菌的体外抗菌效应研究,指导临床正确选择并合理使用抗菌药物。方法 收集2012—2016年期间分离自温州医科大学附属第三医院临床样本中对碳青霉烯类耐药的肺炎克雷伯菌109株,采用Vitek-2 compact和Vitek MS进行菌种鉴定及药敏试验;采用聚合酶链反应(PCR)法检测菌株的KPC酶耐药基因并采用琼脂稀释法药敏试验分别检测替加环素、多黏菌素等药物的单药最低抑菌浓度(minimal inhibitory concentration, MIC)值。结果 所分离的109株均为产KPC-2肺炎克雷伯菌,经过Vitek-2 compact和KB法试验可知109株肺炎克雷伯菌对阿米卡星的耐药率为82.57%,对亚胺培南、美罗培南等其他药物的耐药率均为100%。琼脂稀释法药敏结果显示109株肺炎克雷伯菌对亚胺培南、美罗培南耐药率均为100%;对阿米卡星耐药率为89%;对替加环素的敏感率为65.1%,29.4%中介;对多黏菌素的敏感率为100%。结论 产KPC-2型肺炎克雷伯菌对多黏菌素和替加环素较为敏感,可作为临床用药参考。     

嗜麦芽寡养单胞菌对黏菌素的耐药特征及体外联合抗菌活性研究

目的 研究黏菌素分别与氯霉素、左氧氟沙星、头孢他啶、米诺环素及复方磺胺甲噁唑联用对多重耐药嗜麦芽寡养单胞菌的体外抗菌活性。方法 收集2015—2018年分离自温州医科大学附属第一医院的431株嗜麦芽寡养单胞菌;采用微量肉汤稀释法检测黏菌素对嗜麦芽寡养单胞菌的最低抑菌浓度(minimal inhibitory concentration, MIC),计算逐年耐药率;通过棋盘法和微量肉汤稀释法检测黏菌素分别与氯霉素、左氧氟沙星、头孢他啶、米诺环素与复方磺胺甲噁唑联用及各自单用时对多重耐药嗜麦芽寡养单胞菌的MIC值,并通过计算部分抑菌浓度指数(FICI)评价联合抑菌效果。结果 2015—2018年间,我院嗜麦芽寡养单胞菌对黏菌素的耐药率呈现逐年上升趋势;黏菌素与氯霉素、左氧氟沙星及米诺环素联用后均表现为协同或相加作用;与头孢他啶及复方磺胺甲噁唑联用都存在无关作用,未发现拮抗作用。结论     

嗜麦芽寡养单胞菌对黏菌素的耐药特征及体外联合抗菌活性研究

目的 研究黏菌素分别与氯霉素、左氧氟沙星、头孢他啶、米诺环素及复方磺胺甲噁唑联用对多重耐药嗜麦芽寡养单胞菌的体外抗菌活性。方法 收集2015—2018年分离自温州医科大学附属第一医院的431株嗜麦芽寡养单胞菌;采用微量肉汤稀释法检测黏菌素对嗜麦芽寡养单胞菌的最低抑菌浓度(minimal inhibitory concentration, MIC),计算逐年耐药率;通过棋盘法和微量肉汤稀释法检测黏菌素分别与氯霉素、左氧氟沙星、头孢他啶、米诺环素与复方磺胺甲噁唑联用及各自单用时对多重耐药嗜麦芽寡养单胞菌的MIC值,并通过计算部分抑菌浓度指数(FICI)评价联合抑菌效果。结果 2015—2018年间,我院嗜麦芽寡养单胞菌对黏菌素的耐药率呈现逐年上升趋势;黏菌素与氯霉素、左氧氟沙星及米诺环素联用后均表现为协同或相加作用;与头孢他啶及复方磺胺甲噁唑联用都存在无关作用,未发现拮抗作用。结论 黏菌素与氯霉素、左氧氟沙星、米诺环素联合对多重耐药嗜麦芽寡养单胞菌具有较好的体外抗菌活性,将对临床联合用药治疗多重耐药嗜麦芽寡养单胞菌的选择更具指导意义。     

莫西沙星对耐药葡萄球菌生物膜药效学研究

目的：研究莫西沙星对4株临床耐药葡萄球菌生物膜的体外药效学。方法：采用微量肉汤稀释法测定莫西沙星的最低抑菌浓度（Minimal inhibitory concentration,MIC）、最低抑制生物被膜浓度（Minimal biofilm inhibitory concentration,MBIC）和最低摧毁生物被膜浓度（minimal biofilm eradication concentration,MBEC）;测定莫西沙星对细菌生物膜形成量以及存活菌的影响;采用微量稀释棋盘法测定莫西沙星与局部用药瑞他帕林的联合抗生物膜效果。结果：莫西沙星在16~256mg/L的范围内可完全摧毁细菌生物膜;2 × MIC显著降低生物被膜的形成量;100 × MIC可显著降低生物被膜存活菌数;与瑞他帕林的联合抗生物膜分数（fractional biofilm inhibitory concentration,FBIC）均小于1.0。结论：莫西沙星对4株临床耐药葡萄球菌生物被膜具有抑制和摧毁作用,而且与局部用药瑞他帕林具有协同作用。     

替加环素对多重耐药鲍曼不动杆菌adeB基因表达水平的影响及临床意义北大核心CSCD

目的探讨替加环素对多重耐药鲍曼不动杆菌ade B基因表达水平的影响及临床意义。方法选取临床分离的60株多重耐药鲍曼不动杆菌(MDR-AB),从中筛选出对环丙沙星(CIP)外排泵编码基因呈阳性的实验菌株15株,用琼脂二倍稀释法检测替加环素作用前后对15株实验菌株的最小抑菌浓度(MIC),对替加环素作用前后MIC变化相对较大的菌株基因进行测序并对比分析。结果替加环素对MDR-AB的MIC最低为2 mg·m L-1;质量浓度为0,0.62 mg·m L-1替加环素同质量浓度为1.25,2.5 mg·m L-1环丙沙星对MDR-AB的MIC值差异有统计学意义(P<0.05)。替加环素作用后,ade B基因片段有15个碱基突变,5处氨基酸发生替换,还有4个突变氨基酸未发生替换。替加环素作用前后ade B基因表达水平分别为(2.51±0.72),(0.87±0.21),差异有统计学意义(P<0.05)。结论替加环素可抑制MDR-AB外排泵ade B基因表达,降低ade B表达水平,环丙沙星联合替加环素治疗MDR-AB可能提高临床疗效。     

多黏菌素B药敏条在肠杆菌科药敏检测中应用研究

目的 使用微量稀释法和E-test法分析肠杆菌科细菌对多黏菌素B体外敏感性，评估E-test法多黏菌素B药敏条在临床药敏检测中的价值。方法 收集了887株临床肠杆菌科细菌，同时采用E-test法和微量稀释法进行体外药敏试验，考察两种方法的一致性，针对不一致的进行复测，复测仍不一致者判为不一致，对两种检测方法进行统计学分析。结果 887株菌的平行检测中，842株用药敏条和微量稀释法测定MIC值在|±1|个稀释倍数的范围内，有45株菌的最低抑菌浓度(minimal inhibitory concentration, MIC)值超出了此范围，复测有8株菌的MIC值回到了|±1|个稀释倍数的范围内，即纠正后的EA(essential agreement)%为95.94%(851/887)。肠杆菌科内按照种属分析，EA值最高的为沙门菌属的97.09%(100/103)，EA最低的为其他属肠杆菌的94.03%(126/134)，统计学分析，不同属间无显著性差异。结论 多黏菌素B药敏条用于肠杆菌科细菌检测药敏检测所测定的MIC值判定结果准确可靠，与CLSI推荐的微量稀释法的符合率在94%以上，能满足临床和实验室药敏检测需求。     

依替米星联合美罗培南、哌拉西林他唑巴坦对多重耐药铜绿假单胞菌突变选择窗的研究

目的:探讨依替米星(ETM)与美罗培南(MEM)、哌拉西林他唑巴坦(TZP)体外联合对多重耐药铜绿假单胞菌(MDRP)耐药突变选择窗的影响,为降低细菌耐药的产生提供依据。方法:分别采用棋盘法、琼脂平板稀释法测定24株临床分离MDRP的最低抑菌浓度(MIC)和防耐药突变浓度(MPC),并计算相应的部分抑菌浓度(FIC)指数及选择指数(SI)。结果:ETM联用MEM、TZP后,对标准菌株的MIC、MPC、SI值较单用时明显降低,并表现为协同作用。ETM+MEM与ETM+TZP对24株MDRP有协同抗菌作用的分别占37.50%、33.33%,相加作用分别占58.33%、41.67%,无关作用分别占4.17%、25.00%,均无拮抗作用。ETM、MEM、TZP单用对24株MDRP的SI分别为8~32、8~32、8~32,ETM分别与MEM、TZP联合后各自单药的SI分别为ETM+MEM(ETM-1~8,MEM-1~8)、ETM+TZP(ETM-1~8,TZP-1~16)。结论:ETM与MEM、TZP联合后,可降低各药对铜绿假单胞菌的MPC值,缩小MSW,有利于防止细菌耐药突变体的产生。     

The combination effects of antibacterial agents against clinical isolated multiple-drug resistant Pseudomonas aeruginosa

The effectiveness of antibacterial agents against 70 strains of clinically isolated multiple-drug resistant Pseudomonas aeruginosa (MDRP) was measured by the micro dilution method. Fifty of all strains (71%) produced metallo-beta-lactamase and the IMP-1 gene was detected by polymerase chain reaction (PCR). The MIC90 (the minimum inhibitory concentration of an antibiotic necessary to inhibit the growth of 90% of bacterial strains) values of biapenem (BIPM), meropenem (MEPM), tazobactam/piperacillin (TAZ/PIPC), sulbactam/ cefoperazone (SBT/CPZ), cefepime (CFPM), ciprofloxacin (CPFX), pazufloxacin (PZFX), amikacin (AMK) and aztreonam (AZT) were found to be 265, 512, 256, 512, 512, 64, 128, 128 and 128 microg/mL, respectively. The in vitro combination effects of antibacterial agents were examined against 62 strains of MDRP and the synergy or additive effects were evaluated by fractional inhibitory concentration (FIC) index calculated by the checkerboard method. The combination of AMK and AZT showed synergy effects on 15/59 (25.4%) strains of MDRP. The synergy and additive effects on the MDRP strains were also found by the other antibacterial agents combination such as TAZ/PIPC and AMK, CFPM and AMK, and SBT/CPZ and AZT. These results suggested the necessity of further investigation of clinical usefulness.     

硝酸镓对耐碳青霉烯肺炎克雷伯菌生长及其生物膜的抑制作用

目的 检测新型抗菌剂硝酸镓(gallium nitrate, GaN)对耐碳青霉烯肺炎克雷伯菌(carbapenem-resistant Klebsiella pneumonia, CRKP)生长及其生物膜(biofilm, BF)的抑制作用。方法 收集2015-2016年安徽省地区34家医院临床分离的CRKP共82株，采用VITEK-2 Compact全自动微生物分析系统进行细菌鉴定，琼脂倍比稀释法进行药敏试验；微量肉汤稀释法测定GaN对CRKP的最低抑菌浓度(minimal inhibitory concentration, MIC)；96孔板结晶紫染色法检测82株实验菌株BF的A570值，筛选出BF强阳性菌株；检测1/2MIC的GaN对BF强阳性菌株的BF形成的抑制作用。比较GaN与庆大霉素(gentamicin, GEN)分别对CRKP的BF形成的抑制作用。激光共聚焦扫描显微镜观察GaN作用下的菌株生物膜三维立体结构。结果 临床分离的CRKP呈多重耐药。GaN对82株CRKP的MIC范围为0.0625~2μmol/mL，其中MIC50、MIC90分别为0.25和0.5μmol/mL。BF强阳性菌株所占比例为37.8%(31/82)。三维立体图片显示CRKP在不同GaN浓度作用下BF形成的量不同。1/2MIC的GaN可显著抑制CRKP的BF形成。GaN对CRKP的BF形成的抑制作用优于GEN。结论 GaN对CRKP生长及其BF有明显抑制作用。     

Antibacterial activities and PK/PD parameters of aminoglycosides against recent clinical isolates of gram-negative rods

We examined antibacterial activities and PK/PD parameters of six kinds of aminoglycosides against seven bacterial species of clinical isolates in 2001. Aminoglycoseides examined were gentamicin (GM), dibekacin (DKB), tobramycin (TOB), amikacin (AMK), netilmicin (NTL), and isepamicin (ISP), and bacterial isolates used were each 50 strains of Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Citrobacter freundii, Proteus spp., Serratia marcescens and Pseudomonas aeruginosa. All aminoglycosides showed good activities with low MICs against 6 species of Enterobacteriacea except S. marcescens. Eight strains (3.2%) among them were resistant to one or more aminoglycosides. Resistance to multiple aminoglycosides were detected in 16 strains (32%) of S. marcescens, among which 13 strains were resistant to AMK but susceptible to ISP. Three (6%) strains of P. aeruginosa were resistant to multiple drugs, one of which was resistant to all six aminoglycosides, and others were moderately susceptible to AMK and ISP, and susceptible to GM, AMK and ISP. Using a ratio of peak serum concentration to MIC90 (Cmax/MIC90) or a ratio of area under the curve to MIC90 (AUC/MIC90) as a pharmacokinetic and pharmacodynamic (PK/PD) parameter, we estimated the efficacy of the drug. An excellent effect of ISP, which was injected intramuscularly or intravenously at a dose of 400 mg, was expected for strains of Enterobacteriacea except S. marcescens. The Cmax/MIC90 ratios for S. marcescens were comparably higher in GM and ISP and that for P. aeruginosa were rather high in TOB when compared to other aminoglycosides. Another PK/PD parameter, AUC/MIC90 ratio, was high enough in NTL and ISP for Enterobacteriacea, suggesting good efficacy of these drugs. The (AUC/MIC90) ratios for S. marcescens were comparably high in GM and ISP, and that for P. aeruginosa were high in TOB, DKB, and ISP.     

The resistance of recent clinical isolates against isepamycin, other aminoglycosides and injectable beta-lactams

Clinical isolates collected from clinical facilities across Japan in 1998 were tested against five aminoglycosides and three beta-lactams. The resistance of 50 strains each of methicillin sensitive Staphylococcus aureus, methicillin resistant Staphylococcus aureus (MRSA), Staphylococcus epidermidis, Escherichia coli, Citrobacter freundii, Klebsiella pneumoniae, Enterobacter sp., Serratia sp., Pseudomonas aeruginosa and Proteus sp. (P. mirabilis 25 strains and P. vulgaris 25 strains) to the aminoglycosides isepamicin (ISP), amikacin (AMK), gentamicin, tobramycin and dibekacin, and to the beta-lactams imipenem, ceftazidime and piperacillin (all three known to be effective against P. aeruginosa) were investigated using a micro liquid dilution method with the following results: 1. ISP was effective against all strains except for 14% of MRSA, 2% of Proteus sp., and 4% of P. aeruginosa. 2. Six strains of MRSA were resistant to all eight drugs; however, in these cases ISP exhibited a relatively low minimum inhibitory concentration (MIC) compared to the other compounds. 3. Four strains of MRSA were resistant to all drugs except ISP. MRSA was the only isolate to demonstrate a resistance to seven or more drugs. 4. Twenty-one strains of MRSA and 1 strain of P. aeruginosa were resistant to six drugs; however, all of these were susceptible to both ISP and AMK. 5. Against all strains tested, ISP generally exhibited a lower MIC compared to AMK. These results suggest that, even ten years after its entering the market, ISP is still an aminoglycoside having a high anti-bacterial activity against a wide range of clinical isolates.     

安徽地区耐碳青霉烯鲍曼不动杆菌分子流行病学研究

目的 研究安徽地区耐碳青霉烯鲍曼不动杆菌(carbapenem-resistant Acinetobacter baumannii, CRAB)的耐药性、碳青 霉烯酶基因型及分子流行病学研究,为控制医院感染提供理论依据。方法 收集安徽地区43家三级医院2015年9月和2016年9月 CRAB非重复菌株312株,从中挑选出两年内均有CRAB的医院12家,菌株147株。对挑选出的147株CRAB采用琼脂稀释法测定 16种抗菌药物的最低抑菌浓度(minimal inhibitory concentration, MIC);PCR扩增碳青霉烯酶耐药基因;采用多位点序列分型技 术(multilocus sequence typing, MLST)进行分子分型,使用eBURST软件分析菌株亲缘性。结果 147株临床分离的CRAB均为多 重耐药菌株,对16种抗菌药物耐药性均较高,其中对替加环素和米诺环素耐药性较低,分别为9.27%和54.26%。147株CRAB中 有134株携带blaOXA-23基因,1株携带blaOXA-24 基因,4株携带blaIMP基因,全部携带blaOXA-51基因,其余碳青霉稀类耐药基因均未检 出。147株CRAB分为26个ST基因型,其中13个ST分型属于CC92克隆群,ST195有48株,为安徽地区最常见ST型,另外新发现 15个ST基因型。结论 本研究中安徽地区147株CRAB对临床常用抗生素耐药性严重,除替加环素和米诺环素外,耐药率均在 76%以上。blaOXA-51是位于染色体上的固有基因,检出率为100%;blaOXA-23 是安徽地区最主要的碳青霉烯酶基因型,检出率为 91.15%。CC92为安徽地区最主要克隆群,与世界流行克隆群一致。本研究所测147株菌株间存在进化间关系,提示可能存在水 平传播趋势。