我院铜绿假单胞菌的耐药性和耐药基因的检测与分析

发布时间：2018-01-04 02:00

  本文关键词：我院铜绿假单胞菌的耐药性和耐药基因的检测与分析　出处：《天津医科大学》2015年硕士论文　论文类型：学位论文

  更多相关文章： 铜绿假单胞菌 β-内酰胺酶 金属酶 外膜蛋白OprD2 耐药的基因

【摘要】：目的:分析天津市北辰区中医院PA(Pseudomonas aeruginosa,铜绿假单胞菌)的检出情况和耐药性,为指导临床控制铜绿假单胞菌感染、合理使用抗菌药及延缓细菌耐药产生提供参考;另外,为了了解本院铜绿假单胞菌的耐药机制,指导临床合理用药,本论文从β-内酰胺酶编码基因种类、存在状况和外膜蛋白Opr D2基因缺失状况方面分别进行了初步研究。方法:1.采集我院住院患者送检标本分离的铜绿假单胞菌共58株,鉴定菌株的室验采用ATB Expression细菌鉴定系统,来源于法国梅里埃公司;药敏试验采用ATB Expression细菌药敏板条,以及琼脂纸片扩散(K-B)法进行细菌药物敏感试验。2用PCR(polymerase chain reaction,聚合酶链反应)法检测17种铜绿假单胞菌相关耐药基因。3对铜绿假单胞菌其中几种耐药基因的PCR扩增产物进行了测序,并将测序结果与Gen Bank登录的结果进行比对。结果1.通过对18种抗生素做药敏试验,58株铜绿假单胞菌耐药率高的有氨苄西林+舒巴坦100%、头孢克洛95.4%、头孢丙烯95.4%、头孢克肟94.4%、头孢美唑94.3%,耐药率较高的是替卡西林/克拉维酸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、头孢曲松、庆大霉素、诺氟沙星、左氧氟沙星、氨曲南、头孢哌酮/舒巴坦、阿米卡星的耐药率为36.6%、32.1%、31.1%、26%、22.9%、17.4%、16.5%、15.5%、15.5%、12.8%、7.33%,其中亚胺培南的耐药率为10%。2.58株铜绿假单胞菌中β-内酰胺酶编码基因及检出率: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%);金属β-内酰胺酶基因及检出率为:ⅥM(25.9%)、IMP(19%)、SPM(6.8%)、GIM(0%)、SIM(0%);质粒Amp C酶基因DHA的检出率是10.3%;膜孔蛋白Opr D2基因的缺失率为48.3%。3.VEB、SHV、TEM和DHA4个基因的核苷酸序列与Genebank中参比基因序列同源性均为100%,未发现基因变异现象。结论:1.本院铜绿假单胞菌对氨苄西林+舒巴坦的耐药率最高为100%,对于氨苄西林铜绿假单胞菌是天然耐药,即使加了舒巴坦酶抑制剂,我院统计耐药率仍然是百分之百;其次是头孢克洛为95.4%、头孢丙烯为95.4%,头孢克肟为94.4%,头孢美唑也达到了94.3%,对于这些耐药率超过75%的抗菌药物,应暂停该类抗菌药物的临床应用,根据追踪细菌耐药监测结果,再决定是否恢复其临床应用。耐药率超过50%的抗菌药物为替卡西林/克拉维酸64.6%,应参照药敏试验结果选用抗生素;耐药率超过40%的抗菌药物,应慎重经验用药;耐药率超过30%的抗菌药物有磷霉素36.6%、依替米星32.1%、阿洛西林31.1%,应及时将预警信息通报本院医护人员;低于30%的抗菌药物有哌拉西林/他唑巴坦26.0%、头孢曲松22.9%、庆大霉素17.4%、诺氟沙星16.5%、氨曲南15.5%、头孢哌酮/舒巴坦12.8%、亚胺培南10.0%,可能会更合理地成为治疗混合感染的二线药;氟喹诺酮类抗菌药物诺氟沙星耐药率16.5%和左氧氟沙星耐药率15.5%,虽然耐药率较低,但此类药物,如果长期使用也可能发生耐药,所以测试重复分离菌株可能是必要的;单环类抗生素的代表氨曲南(15.5%)有抗PA活性,但较弱,而且临床反应效果不好,疗效有限,仅适用于轻、中度感染;阿米卡星的耐药率最低为7.33%,一般不单独用于治疗,但可以用于联合治疗。2.β-内酰胺酶编码基因的检出率比较高(79%),阳性率比较高的基因是:OXA-1(32.7%)、VIM(25.9%)、CTX-M-1(24.1%);膜孔蛋白Opr D2基因的缺失率比较高达48.3%,说明或许是很多种耐药的基因共同作用产生了PA对β-内酰胺类抗菌药的多重耐药性,此菌具有非常复杂的耐药机制。3 VEB、SHV、TEM、和DHA4个基因的核苷酸序列与Genebank中参比基因序列同源性均为100%,未曾发现基因的变异现象。4铜绿假单胞菌(PA)的感染现状和耐药情况不容乐观,需要进一步加强监测其耐药性及研究其耐药基因的分子流行病学,针对于本院及本地区感染制定更科学的治疗方案,防止耐药菌株播散,减少院内感染的发生,提高铜绿假单胞菌临床感染的治愈率。
[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�閾滅豢鍋囧崟鑳炶弻涓�尾-鍐呴叞鑳洪叾缂栫爜鍩哄洜鍙婃��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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