宁夏地区嗜麦芽窄食单胞菌临床耐药分布及氟喹诺酮耐药基因研究
发布时间：2019-05-23 14:16
【摘要】：目的对嗜麦芽窄食单胞菌感染的临床分布、抗菌药物的耐药状况及耐药基因进行研究,为临床治疗提供有力依据;用脉冲场凝胶电泳技术(PFGE)对嗜麦芽窄食单胞菌进行同源性分析,探讨该菌的医院感染特征。方法收集2010—2012年三年宁夏医科大学总医院临床常规检出的嗜麦芽窄食单胞菌,采用全自动微生物鉴定仪对细菌进行鉴定,并用K-B琼脂纸片扩散法进行药敏试验;筛选2012年全年114株嗜麦芽窄食单胞菌,采用琼脂二倍稀释法测定7种抗菌药物对细菌的最低抑菌浓度(MIC)值,同批测定诺氟沙星、环丙沙星、氧氟沙星加入外排泵抑制剂(利血平)后抗菌药物的MIC值的变化情况;应用聚合酶链反应(PCR)检测嗜麦芽窄食单胞菌中氟喹诺酮类耐药基因的存在情况;应用实时荧光定量PCR法检测外排泵sme D和sme F基因m RNA的相对表达水平;测序后进行Blast比对序列;应用脉冲场凝胶电泳进行同源性分析。结果2010—2012年三年共分离得到嗜麦芽窄食单胞菌426株,占非发酵菌的10.1%(426/4225);114株嗜麦芽窄食单胞菌对诺氟沙星、环丙沙星和氧氟沙星的耐药率分别为32.4%、21.9%和13.2%。加入泵抑制剂后共筛选得到19株外排泵阳性株。筛选得到的2株外排泵阳性的耐药株均有sme D和sme F的高表达。114株嗜麦芽窄食单胞菌中gyr A,par C,sme D,sme E,sme F基因的阳性率均为100%,smqnr基因的阳性率为25.4%。5株菌的gyr A基因有3株菌的第51位氨基酸发生了由谷氨酸突变为赖氨酸;5株菌的par C基因中有4株菌的第37位的氨基酸发生了由甘氨酸突变为精氨酸。发生gyr A或par C基因突变的耐药与敏感组之间的差异无显著性(P0.05)。对29株嗜麦芽窄食单胞菌进行PFGE同源性分析,呈现散在分布。结论1.宁夏医科大学总医院的嗜麦芽窄食单胞菌主要分布在ICU、呼吸科、神经外科以及儿科,以痰、咽拭子、导管和排泄物中检出率最高。2.在嗜麦芽窄食单胞菌中smqnr基因介导氟喹诺酮的低水平耐药且与外排泵基因共同介导耐药。3.通过PFGE技术得出本院嗜麦芽窄食单胞菌并未出现同一克隆株的暴发,呈现散在分布。
[Abstract]:Objective to study the clinical distribution, antibiotic resistance and drug resistance genes of Stenococcus maltophilia infection, so as to provide a powerful basis for clinical treatment. The homology of Stenococcus maltophilia was analyzed by pulse field gel electrophoresis (PFGE), and the characteristics of hospital infection of Stenococcus maltophilia were discussed. Methods from 2010 to 2012, Stenococcus maltophilia was collected from the General Hospital of Ningxia Medical University. The bacteria were identified by automatic microbial identification instrument, and the drug sensitivity test was carried out by K / B Agar disk diffusion method. 114 strains of Stenococcus maltophilia were screened for the whole year of 2012. The minimum inhibitory concentration (MIC) of 7 antibiotics against bacteria was determined by Agar dilution method, and norfloxacin and ciprofloxacin were determined in the same batch. The changes of MIC of antibiotics after ofloxacin was added to efflux pump inhibitor (reserpine). The presence of fluoroquinolones resistance genes in Stenococcus maltophilia was detected by polymerase chain reaction (PCR), and the relative expression of sme D and sme F gene m RNA in efflux pump was detected by real-time fluorescence quantitative PCR. The sequence was compared by Blast after sequencing, and the homology was analyzed by pulse field gel electrophoresis. Results A total of 426 strains of Stenococcus maltophilia were isolated from 2010 to 2012, accounting for 10.1% (426 鈮，
本文编号：2483969
[Abstract]:Objective to study the clinical distribution, antibiotic resistance and drug resistance genes of Stenococcus maltophilia infection, so as to provide a powerful basis for clinical treatment. The homology of Stenococcus maltophilia was analyzed by pulse field gel electrophoresis (PFGE), and the characteristics of hospital infection of Stenococcus maltophilia were discussed. Methods from 2010 to 2012, Stenococcus maltophilia was collected from the General Hospital of Ningxia Medical University. The bacteria were identified by automatic microbial identification instrument, and the drug sensitivity test was carried out by K / B Agar disk diffusion method. 114 strains of Stenococcus maltophilia were screened for the whole year of 2012. The minimum inhibitory concentration (MIC) of 7 antibiotics against bacteria was determined by Agar dilution method, and norfloxacin and ciprofloxacin were determined in the same batch. The changes of MIC of antibiotics after ofloxacin was added to efflux pump inhibitor (reserpine). The presence of fluoroquinolones resistance genes in Stenococcus maltophilia was detected by polymerase chain reaction (PCR), and the relative expression of sme D and sme F gene m RNA in efflux pump was detected by real-time fluorescence quantitative PCR. The sequence was compared by Blast after sequencing, and the homology was analyzed by pulse field gel electrophoresis. Results A total of 426 strains of Stenococcus maltophilia were isolated from 2010 to 2012, accounting for 10.1% (426 鈮，
本文编号：2483969
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