保定市多重耐药铜绿假单胞菌氨基糖苷类修饰酶基因的检测及分析

发布时间：2018-01-29 05:28

  本文关键词： 铜绿假单胞菌 耐药基因 氨基糖苷类修饰酶 聚合酶链反应 保定地区　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:1掌握保定市临床分离铜绿假单胞菌的耐药性情况。2掌握多重耐药铜绿假单胞菌氨基糖苷类耐药基因的分布情况,为指导临床合理使用抗生素、减缓细菌耐药产生、有效预防和控制院内感染提供依据。方法:1菌株来源从2016年1月至2016年8月,收集保定市第一中心医院临床标本中分离培养出的不重复之铜绿假单胞菌,按照《全国临床检验操作规程》第三版的标准,用VITEK-2全自动微生物鉴定仪鉴定菌种。2鉴定及药敏试验对铜绿假单胞菌菌种进行纯化,用vitek-2全自动微生物鉴定仪鉴定菌种,而后采用K-B法测定其对阿米卡星等20种抗菌药物的敏感性。3耐药基因检测选取多重耐药铜绿假单胞菌,使用Maxwell?16 Cell LEV DNA Purification Kit试剂盒提取DNA,利用耐药基因检测试剂盒进行聚合酶链反应(PCR),扩增(aac(3)-Ⅰ、aac(3)-Ⅱ、aac(6′)-Ⅰb、aac(6′)-Ⅱ、ant(3″)-Ⅰ、ant(2″)-Ⅰ)6种氨基糖苷类耐药基因。PCR产物用2.0%琼脂糖凝胶电泳,出现与阳性对照分子相当的目的条带判断为阳性,照相后保存影像。结果:1铜绿假单胞菌药敏试验结果对临床分离出的142株铜绿假单胞菌进行了20种抗生素药敏试验,结果显示:耐药率最高的抗生素分别为头孢唑林(97.1%)、头孢曲松(95.7%)、头孢呋辛酯(95.7%)、头孢呋辛钠(95.1%)、氨苄西林(95.0%)、氨苄西林+舒巴坦(94.3%)、头孢替坦(94.3%)、复方新诺明(92.9%)、头孢哌酮(54.2%)。敏感性最高的抗生素分别为阿米卡星(78.1%)、妥布霉素(71.1%)、哌拉西林+他唑巴坦(64.0%)、头孢他啶(61.95%)、哌拉西林(56.3%)、美罗培南(56.3%)、左氧氟沙星(53.5%)、环丙沙星(52.8%)、庆大霉素(44.3%)、亚胺培南(47.1%)。2氨基糖苷类耐药基因检测结果对32株多重耐药铜绿假单胞菌进行耐药基因检测,检测出aac(3)-Ⅱ、aac(6′)-Ⅰb和ant(3″)-Ⅰ氨基糖苷类耐药基因,阳性率分别为(12.5%)、(18.7%)和(9.3%),未检测到aac(3)-Ⅰ、aac(6′)-Ⅱ、ant(2″)-Ⅰ基因。结论:1 2016年保定市分离出的铜绿假单胞菌对头孢唑林、头孢曲松、头孢呋辛酯、头孢呋辛钠、氨苄西林、氨苄西林/舒巴坦、头孢替坦和复方新诺明8种抗生素有很高的耐药率。2阿米卡星、妥布霉素和庆大霉素三种氨基糖苷类药物对铜绿假单胞菌有较高的敏感性,分别为78.1%、71.1%、44.3%,建议临床优先考虑使用。3检测出的多重耐药铜绿假单胞菌氨基糖苷类耐药基因以aac(3)-Ⅱ、aac(6′)-Ⅰb和ant(3″)-Ⅰ为主,未检测到aac(3)-Ⅰ、ant(2″)-Ⅰ、aac(6′)-Ⅱ基因,说明铜绿假单胞菌产生的多重耐药性与氨基糖苷类修饰酶基因的存在有密切关系。4医院应加强铜绿假单胞菌耐药性和耐药基因的监测,指导临床科学有效地使用抗生素。
[Abstract]:Objective to understand the drug resistance of Pseudomonas aeruginosa isolated in Baoding City. To control the distribution of aminoglycoside resistance genes in multidrug resistant Pseudomonas aeruginosa and to guide the rational use of antibiotics. In order to prevent and control nosocomial infection effectively, the bacteria resistance was slowed down. Methods the source of strain: 1 was from January 2016 to August 2016. The non-repeated Pseudomonas aeruginosa isolated from clinical specimens of the first central hospital of Baoding City was collected according to the standards of the third edition of the National procedure for Clinical examination. The strains of Pseudomonas aeruginosa were purified by VITEK-2 automatic microbiological identification instrument and drug sensitivity test, and identified by vitek-2 automatic microbiological identification instrument. Then K-B method was used to determine its sensitivity to 20 antimicrobial agents such as amikacin. 3. 3. Multidrug resistant Pseudomonas aeruginosa was selected and Maxwellwell was used in the detection of multidrug resistant Pseudomonas aeruginosa. 16 Cell LEV DNA Purification Kit kit was used to extract DNA and polymerase chain reaction (PCR) was used to detect drug resistance gene. The results showed that: 1. Aacanine 3 "- 鈪，

本文编号：1472657