保定市铜绿假单胞菌耐药性分析以及耐亚胺培南菌株基因型检测

发布时间：2018-05-09 09:55

本文选题：铜绿假单胞菌 + 耐药基因　；参考：《河北医科大学》2017年硕士论文

【摘要】：目的:1掌握保定市铜绿假单胞菌耐药情况及特点,指导临床医师合理使用抗生素,延缓细菌耐药产生、有效预防及控制院内感染。2选取耐亚胺培南铜绿假单胞菌,检测β-内酰胺酶编码基因种类和外膜蛋白Opr D2基因缺失状况,探讨铜绿假单胞菌对亚胺培南药物的耐药机制。方法:1标本来源收集保定市第一中心医院住院患者2016年7月-2017年1月送检标本分离的铜绿假单胞菌,根据《全国临床检验操作规程》第3版的要求,用羊血培养基对送检标本进行细菌分离和培养,鉴定菌株采用法国梅里埃公司VITEK-2全自动微生物鉴定仪。2药敏实验将铜绿假单胞菌纯化后进行耐药性检测,采用VITEK-2全自动微生物鉴定仪和K-B法测定其对氨苄西林、亚胺培南等23种常用抗菌药物的敏感性,挑选出耐亚胺培南的铜绿假单胞菌株。3耐药基因检测使用Maxwell?16 Cell LEV DNA Purification Kit试剂盒提取DNA,用聚合酶链反应(PCR)法扩增耐亚胺培南菌株β-内酰胺酶基因和外膜孔道蛋白D2基因。产物经2.0%琼脂糖凝胶电泳,凝胶成像仪观察电泳结果,与标准量DNAMarker相比较,对PCR产物的电泳结果进行分析和摄像,出现与目的基因片段分子相当的条带判为阳性,照相保存影像。结果:1铜绿假单胞菌对常用抗菌药物的耐药情况测定了2016年7月到2017年1月临床分离的122株铜绿假单胞菌对23种抗生素的耐药性,耐药率结果为:氨苄西林98.36%、阿米卡星10.66%、环丙沙星25.41%、庆大霉素16.39%、妥布霉素15.57%、复方新诺明96.72%、氨苄西林/舒巴坦96.72%、头孢他啶17.21%、亚胺培南40.98%、哌拉西林18.03%、头孢唑林97.54%、头孢吡肟20.49%、左氧氟沙星22.95%、美罗培南23.77%、头孢替坦95.9%、哌拉西林+他唑巴坦(常规用量)13.11%、头孢曲松98.36%、头孢哌酮36.07%、头孢哌酮/舒巴坦15.57%、头孢呋辛酯95.9%、头孢呋辛钠95.9%、比阿培南23.77%,呋喃妥因只用于尿液标本的药敏试验,耐药率100%。2β-内酰胺酶编码基因检出情况从50株耐亚胺培南的铜绿假单胞菌检出β-内酰胺酶编码基因TEM(3株)、PER(3株)、VEB(1株)、GES(2株)、OXA-2(2株)、OXA-10(3株)、CTX-M-1(3株)。3金属β-内酰胺酶基因检出情况从50株耐亚胺培南的铜绿假单胞菌检出金属β-内酰胺酶基因VIM(9株)、IMP(11株)、SIM(0株)、SPM(5株)、GIM(0株)。4膜孔蛋白Opr D2基因的缺失情况从50株耐亚胺培南的铜绿假单胞菌检测出孔蛋白Opr D2基因缺失31株,缺失率为62%。结论:1保定市铜绿假单胞菌对头孢呋辛酯、头孢呋辛钠、头孢替坦、头孢曲松、氨苄西林、复方新诺明等抗生素有很高的耐药率。2铜绿假单胞菌对亚胺培南的耐药率40.98%。3 50株耐亚胺培南的铜绿假单胞菌β-内酰胺酶编码基因检出率为58%(29株),其中有10株同时含有两种或两种以上基因;Opr D2基因缺失率高(62%)。提示铜绿假单胞菌对β-内酰胺类抗生素的耐药是由多种耐药基因共同控制的,该菌具有复杂的耐药机制。4应加强铜绿假单胞菌耐药性的监测及耐药基因的分子流行病学研究,防止耐药菌株播散,提高临床治愈率。
[Abstract]:Objective: 1 to master the drug resistance and characteristics of Pseudomonas aeruginosa in Baoding, to guide the clinicians to use antibiotics reasonably, to delay the production of bacterial resistance, to effectively prevent and control.2 in hospital infection, to select imipenem Pseudomonas aeruginosa, to detect the type of beta lactamase coding gene and the absence of Opr D2 gene in the outer membrane protein, and to explore the Pseudomonas aeruginosa. The mechanism of drug resistance of monomonas to imipenem. Methods: 1 specimen sources were collected to collect Pseudomonas aeruginosa isolated from the hospitalized patients in Baoding First Central Hospital in January -2017 July 2016. According to the requirements of the third edition of the national clinical inspection procedure, the samples were isolated and cultured with the sheep blood culture medium to identify the bacteria. The strain of Pseudomonas aeruginosa was purified by the.2 drug sensitivity test of VITEK-2 automatic microorganism identification instrument in France mereer company. The sensitivity of 23 kinds of commonly used antibiotics, such as ampicillin and imipenem, was determined by VITEK-2 automatic microorganism identification instrument and K-B method, and the Pseudomonas aeruginosa was selected. The strain.3 resistance gene was detected by using Maxwell? 16 Cell LEV DNA Purification Kit kit to extract DNA, and polymerase chain reaction (PCR) method was used to amplify the gene of beta lactamase and outer membrane D2 gene of imipenem resistant strain. The product was observed by 2% agarose gel electrophoresis and gel imaging instrument, compared with the standard quantity DNAMarker. The electrophoretic results of PCR products were analyzed and photographed, and the bands of the target genes were found to be positive, and the images were preserved. Results: 1 the resistance of Pseudomonas aeruginosa to commonly used antibiotics was tested for the resistance of 122 strains of Pseudomonas aeruginosa to 23 antibiotics from July 2016 to January 2017, and the resistance of Pseudomonas aeruginosa to January 2017 was tested. The drug rate was ampicillin 98.36%, Amikacin 10.66%, ciprofloxacin 25.41%, gentamicin 16.39%, tobramycin 15.57%, compound novamethoxine 96.72%, ampicillin / sulbactam 96.72%, ceftazidime 17.21%, imipenem 40.98%, piperacillin 18.03%, cefazolin 97.54%, cefpyoxime 20.49%, levofloxacin 22.95%, meropenem 23.77%, Ceftaetan 95.9%, piperacillin + tazobactam (conventional dosage) 13.11%, ceftriaxone 98.36%, cefoperazone 36.07%, Cefoperazone / sulbactam 15.57%, cefuroxime 95.9%, cefuroxime sodium 95.9%, amperan 23.77%, furadetin only used in urine samples, drug sensitivity test, the resistance rate 100%.2 beta lactamase gene detection from 5 0 strains of Pseudomonas aeruginosa detected beta lactamase encoding gene TEM (3 strains), PER (3 strain), VEB (1 strain), GES (2 strain), OXA-2 (2 strain), OXA-10 (3 strain), CTX-M-1 (3 strain) gene detection of metallo beta lactamase gene from 50 imipenem resistant Pseudomonas aeruginosa gene VIM (9 strain), IMP (11 strain), SIM (0) strain) SPM (5 strains), GIM (0 strains).4 membrane pore protein Opr D2 gene deletion from 50 strains of imipenem resistant Pseudomonas aeruginosa to detect the loss of pore protein Opr D2 gene deletion, the loss rate is 62%. conclusion: 1 Baoding Pseudomonas aeruginosa, cefuroxime sodium, cefuroxime, ceftriaxone, ampicillin, ampicillin, compound new nooxin and other antibiotics The resistance rate of Pseudomonas aeruginosa to imipenem.2 was highly resistant to imipenem 40.98%.3 50 strains of Pseudomonas aeruginosa resistant to Pseudomonas aeruginosa was 58% (29 strains), of which 10 had two or more than two genes, and the Opr D2 gene deletion rate was high (62%). The resistance of raw materials is controlled by multiple resistance genes, which has a complex resistance mechanism.4 should strengthen the monitoring of the resistance of Pseudomonas aeruginosa and the molecular epidemiology of resistance genes to prevent the spread of drug-resistant strains and improve the clinical cure rate.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R440

【参考文献】