本文關(guān)鍵詞：我院銅綠假單胞菌的耐藥性和耐藥基因的檢測與分析　出處：《天津醫(yī)科大學》2015年碩士論文　論文類型：學位論文

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【摘要】：目的:分析天津市北辰區(qū)中醫(yī)院PA(Pseudomonas aeruginosa,銅綠假單胞菌)的檢出情況和耐藥性,為指導(dǎo)臨床控制銅綠假單胞菌感染、合理使用抗菌藥及延緩細菌耐藥產(chǎn)生提供參考;另外,為了了解本院銅綠假單胞菌的耐藥機制,指導(dǎo)臨床合理用藥,本論文從β-內(nèi)酰胺酶編碼基因種類、存在狀況和外膜蛋白Opr D2基因缺失狀況方面分別進行了初步研究。方法:1.采集我院住院患者送檢標本分離的銅綠假單胞菌共58株,鑒定菌株的室驗采用ATB Expression細菌鑒定系統(tǒng),來源于法國梅里埃公司;藥敏試驗采用ATB Expression細菌藥敏板條,以及瓊脂紙片擴散(K-B)法進行細菌藥物敏感試驗。2用PCR(polymerase chain reaction,聚合酶鏈反應(yīng))法檢測17種銅綠假單胞菌相關(guān)耐藥基因。3對銅綠假單胞菌其中幾種耐藥基因的PCR擴增產(chǎn)物進行了測序,并將測序結(jié)果與Gen Bank登錄的結(jié)果進行比對。結(jié)果1.通過對18種抗生素做藥敏試驗,58株銅綠假單胞菌耐藥率高的有氨芐西林+舒巴坦100%、頭孢克洛95.4%、頭孢丙烯95.4%、頭孢克肟94.4%、頭孢美唑94.3%,耐藥率較高的是替卡西林/克拉維酸64.6%,其余抗生素磷霉素、依替米星、阿洛西林、哌拉西林/他唑巴坦、頭孢曲松、慶大霉素、諾氟沙星、左氧氟沙星、氨曲南、頭孢哌酮/舒巴坦、阿米卡星的耐藥率為36.6%、32.1%、31.1%、26%、22.9%、17.4%、16.5%、15.5%、15.5%、12.8%、7.33%,其中亞胺培南的耐藥率為10%。2.58株銅綠假單胞菌中β-內(nèi)酰胺酶編碼基因及檢出率:TEM(15.5%)、TEM-1(10.3%)、VEB(1.7%)、SHV(6.8%)、GES(3.4%)、PER(3.4%)、OXA-1(32.7%)、OXA-2(15.5%)、OXA-10(13.7%)、CTX-M-1(24.1%);金屬β-內(nèi)酰胺酶基因及檢出率為:ⅥM(25.9%)、IMP(19%)、SPM(6.8%)、GIM(0%)、SIM(0%);質(zhì)粒Amp C酶基因DHA的檢出率是10.3%;膜孔蛋白Opr D2基因的缺失率為48.3%。3.VEB、SHV、TEM和DHA4個基因的核苷酸序列與Genebank中參比基因序列同源性均為100%,未發(fā)現(xiàn)基因變異現(xiàn)象。結(jié)論:1.本院銅綠假單胞菌對氨芐西林+舒巴坦的耐藥率最高為100%,對于氨芐西林銅綠假單胞菌是天然耐藥,即使加了舒巴坦酶抑制劑,我院統(tǒng)計耐藥率仍然是百分之百;其次是頭孢克洛為95.4%、頭孢丙烯為95.4%,頭孢克肟為94.4%,頭孢美唑也達到了94.3%,對于這些耐藥率超過75%的抗菌藥物,應(yīng)暫停該類抗菌藥物的臨床應(yīng)用,根據(jù)追蹤細菌耐藥監(jiān)測結(jié)果,再決定是否恢復(fù)其臨床應(yīng)用。耐藥率超過50%的抗菌藥物為替卡西林/克拉維酸64.6%,應(yīng)參照藥敏試驗結(jié)果選用抗生素;耐藥率超過40%的抗菌藥物,應(yīng)慎重經(jīng)驗用藥;耐藥率超過30%的抗菌藥物有磷霉素36.6%、依替米星32.1%、阿洛西林31.1%,應(yīng)及時將預(yù)警信息通報本院醫(yī)護人員;低于30%的抗菌藥物有哌拉西林/他唑巴坦26.0%、頭孢曲松22.9%、慶大霉素17.4%、諾氟沙星16.5%、氨曲南15.5%、頭孢哌酮/舒巴坦12.8%、亞胺培南10.0%,可能會更合理地成為治療混合感染的二線藥;氟喹諾酮類抗菌藥物諾氟沙星耐藥率16.5%和左氧氟沙星耐藥率15.5%,雖然耐藥率較低,但此類藥物,如果長期使用也可能發(fā)生耐藥,所以測試重復(fù)分離菌株可能是必要的;單環(huán)類抗生素的代表氨曲南(15.5%)有抗PA活性,但較弱,而且臨床反應(yīng)效果不好,療效有限,僅適用于輕、中度感染;阿米卡星的耐藥率最低為7.33%,一般不單獨用于治療,但可以用于聯(lián)合治療。2.β-內(nèi)酰胺酶編碼基因的檢出率比較高(79%),陽性率比較高的基因是:OXA-1(32.7%)、VIM(25.9%)、CTX-M-1(24.1%);膜孔蛋白Opr D2基因的缺失率比較高達48.3%,說明或許是很多種耐藥的基因共同作用產(chǎn)生了PA對β-內(nèi)酰胺類抗菌藥的多重耐藥性,此菌具有非常復(fù)雜的耐藥機制。3 VEB、SHV、TEM、和DHA4個基因的核苷酸序列與Genebank中參比基因序列同源性均為100%,未曾發(fā)現(xiàn)基因的變異現(xiàn)象。4銅綠假單胞菌(PA)的感染現(xiàn)狀和耐藥情況不容樂觀,需要進一步加強監(jiān)測其耐藥性及研究其耐藥基因的分子流行病學,針對于本院及本地區(qū)感染制定更科學的治療方案,防止耐藥菌株播散,減少院內(nèi)感染的發(fā)生,提高銅綠假單胞菌臨床感染的治愈率。
[Abstract]:Objective: to analyze the PA of Tianjin Beichen Hospital of Traditional Chinese Medicine (Pseudomonas aeruginosa, Pseudomonas aeruginosa) detection and drug resistance for clinical control of Pseudomonas aeruginosa infection, rational use of antibiotics and antibiotic resistance delayed reference; in addition, in order to understand the mechanism of drug resistance of Pseudomonas aeruginosa, guide clinical rational drug use, this paper from the beta lactamase gene encoding type, existing condition and Opr outer membrane protein D2 gene deletion status were studied. Methods: 1. Pseudomonas aeruginosa collected in our hospital were collected from patients with a total of 58 strains, strain identification experiment using ATB Expression bacterial identification system, from the French bioMerieux company; drug sensitivity test by bacterial susceptibility ATB Expression slab, and Kirby Bauer (K-B) method for bacterial drug sensitivity test by PCR (polymerase chain rea.2 Ction, polymerase chain reaction) method for the detection of drug resistance related 17 Pseudomonas aeruginosa.3 gene was sequenced on Pseudomonas aeruginosa which several resistance genes of PCR amplification products, and the sequencing results with the Gen Bank log results were compared. Results 1. through drug sensitivity test to 18 kinds of antibiotics, 58 strains of resistant P. The high rate of Pseudomonas with ampicillin sulbactam + 100%, cefaclor cefprozil 95.4%, 95.4%, 94.4% cefixime, cefmetazole 94.3%, drug resistance rate is higher in ticarcillin / clavulanic acid 64.6%, other antibiotics fosfomycin, etimicin, azlocillin, piperacillin / tazobactam tazobactam, ceftriaxone, norfloxacin, gentamicin, aztreonam, levofloxacin, Cefoperazone / sulbactam, Amikacin's drug resistance rate was 36.6%, 32.1%, 31.1%, 26%, 22.9%, 17.4%, 16.5%, 15.5%, 15.5%, 12.8%, 7.33%, the imipenem resistance was 10%.2.58 鏍摐緇垮亣鍗曡優(yōu)鑿屼腑尾-鍐呴叞鑳洪叾緙栫爜鍩哄洜鍙婃鍑虹巼:TEM(15.5%),TEM-1(10.3%),VEB(1.7%),SHV(6.8%),GES(3.4%),PER(3.4%),OXA-1(32.7%),OXA-2(15.5%),OXA-10(13.7%),CTX-M-1(24.1%);閲戝睘尾-鍐呴叞鑳洪叾鍩哄洜鍙婃鍑虹巼涓，

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