本文關(guān)鍵詞： 銅綠假單胞菌 耐藥基因 氨基糖苷類修飾酶 聚合酶鏈反應(yīng) 保定地區(qū)　出處：《河北醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:1掌握保定市臨床分離銅綠假單胞菌的耐藥性情況。2掌握多重耐藥銅綠假單胞菌氨基糖苷類耐藥基因的分布情況,為指導(dǎo)臨床合理使用抗生素、減緩細菌耐藥產(chǎn)生、有效預(yù)防和控制院內(nèi)感染提供依據(jù)。方法:1菌株來源從2016年1月至2016年8月,收集保定市第一中心醫(yī)院臨床標(biāo)本中分離培養(yǎng)出的不重復(fù)之銅綠假單胞菌,按照《全國臨床檢驗操作規(guī)程》第三版的標(biāo)準(zhǔn),用VITEK-2全自動微生物鑒定儀鑒定菌種。2鑒定及藥敏試驗對銅綠假單胞菌菌種進行純化,用vitek-2全自動微生物鑒定儀鑒定菌種,而后采用K-B法測定其對阿米卡星等20種抗菌藥物的敏感性。3耐藥基因檢測選取多重耐藥銅綠假單胞菌,使用Maxwell?16 Cell LEV DNA Purification Kit試劑盒提取DNA,利用耐藥基因檢測試劑盒進行聚合酶鏈反應(yīng)(PCR),擴增(aac(3)-Ⅰ、aac(3)-Ⅱ、aac(6′)-Ⅰb、aac(6′)-Ⅱ、ant(3″)-Ⅰ、ant(2″)-Ⅰ)6種氨基糖苷類耐藥基因。PCR產(chǎn)物用2.0%瓊脂糖凝膠電泳,出現(xiàn)與陽性對照分子相當(dāng)?shù)哪康臈l帶判斷為陽性,照相后保存影像。結(jié)果:1銅綠假單胞菌藥敏試驗結(jié)果對臨床分離出的142株銅綠假單胞菌進行了20種抗生素藥敏試驗,結(jié)果顯示:耐藥率最高的抗生素分別為頭孢唑林(97.1%)、頭孢曲松(95.7%)、頭孢呋辛酯(95.7%)、頭孢呋辛鈉(95.1%)、氨芐西林(95.0%)、氨芐西林+舒巴坦(94.3%)、頭孢替坦(94.3%)、復(fù)方新諾明(92.9%)、頭孢哌酮(54.2%)。敏感性最高的抗生素分別為阿米卡星(78.1%)、妥布霉素(71.1%)、哌拉西林+他唑巴坦(64.0%)、頭孢他啶(61.95%)、哌拉西林(56.3%)、美羅培南(56.3%)、左氧氟沙星(53.5%)、環(huán)丙沙星(52.8%)、慶大霉素(44.3%)、亞胺培南(47.1%)。2氨基糖苷類耐藥基因檢測結(jié)果對32株多重耐藥銅綠假單胞菌進行耐藥基因檢測,檢測出aac(3)-Ⅱ、aac(6′)-Ⅰb和ant(3″)-Ⅰ氨基糖苷類耐藥基因,陽性率分別為(12.5%)、(18.7%)和(9.3%),未檢測到aac(3)-Ⅰ、aac(6′)-Ⅱ、ant(2″)-Ⅰ基因。結(jié)論:1 2016年保定市分離出的銅綠假單胞菌對頭孢唑林、頭孢曲松、頭孢呋辛酯、頭孢呋辛鈉、氨芐西林、氨芐西林/舒巴坦、頭孢替坦和復(fù)方新諾明8種抗生素有很高的耐藥率。2阿米卡星、妥布霉素和慶大霉素三種氨基糖苷類藥物對銅綠假單胞菌有較高的敏感性,分別為78.1%、71.1%、44.3%,建議臨床優(yōu)先考慮使用。3檢測出的多重耐藥銅綠假單胞菌氨基糖苷類耐藥基因以aac(3)-Ⅱ、aac(6′)-Ⅰb和ant(3″)-Ⅰ為主,未檢測到aac(3)-Ⅰ、ant(2″)-Ⅰ、aac(6′)-Ⅱ基因,說明銅綠假單胞菌產(chǎn)生的多重耐藥性與氨基糖苷類修飾酶基因的存在有密切關(guān)系。4醫(yī)院應(yīng)加強銅綠假單胞菌耐藥性和耐藥基因的監(jiān)測,指導(dǎo)臨床科學(xué)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，

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