多药耐药铜绿假单胞菌β-内酰胺类耐药相关基因及Ⅰ型整合酶基因研究

Genotypes of β-Lactamases Multi-drug Resistant Genes of Pseudomonas aeruginosa and Type Ⅰ Integrating Enzyme Genes

目的了解临床分离的多药耐药铜绿假单胞菌(MRPA)中β-内酰胺类药物耐药相关基因及Ⅰ型整合酶基因存在状况。方法对34株多药耐药铜绿假单胞菌用K-B纸片扩散法和微量稀释法测定对庆大霉素等16种抗菌药物的敏感性;用PCR法对β-内酰胺类药物耐药相关基因TEM、SHV、OXA-10群、PER、VEB、GES、CARB、IMP、VIM、SPM、GIM、DHA、FOX、MOX、CTX-M9-群、CTX-M-1群和外膜蛋白oprD2基因及Ⅰ型整合酶基因等18种主要耐药基因进行了检测与分析。结果34株MRPA(耐3类抗菌药物的菌株)对阿米卡星、头孢他啶、环丙沙星、头孢哌酮/舒巴坦、头孢吡肟、美罗培南、左氧氟沙星、哌拉西林/他唑巴坦、氨曲南、哌拉西林、妥布霉素的耐药率分别为14.7%、29.4%、61.8%、67.6%、70.6%、73.5%、76.5%、76.5%、79.4%、91.2%、94.1%,对其他抗菌药物耐药率均为100.0%;检出β-内酰胺酶编码基因FOXI、MP、DHA和VIM基因阳性率分别为23.5%、11.8%、11.8%和5.9%,24株(70.6%)外膜通道蛋白oprD2基因缺失,而其他基因均阴性,34株Ⅰ型整合酶基因intⅠ1均阳性。结论临床分离的MRPA携带多种β-内酰胺酶基因,oprD2基因缺失率高,Ⅰ型整合酶基因广泛存在于MRPA。

OBJECTIVE To investigate genes associated with the drug-resistance of β-lactamases antibiotics in Pseudomonas aeruginosa （PAE） and type Ⅰ integrating enzyme gene isolated from clinical patients. METHODS The drug-sensitivity for 16 antibiotics were detected by the disc agar diffusion（DAD） and microdilution. To diteet the related drug-resistenee genes TEM, SHV, OXA-10, PER, VEB, GES, CARB, IMP, VIM, SPM, GIM, DHA, F-OX, MOX, CTX-M9, CTX-M1 and outer membrane protein gene oprD2 and 18 main drug resistance genes of type 1 integrating enzyme gene and others with PCR method. RESULTS Resistant rates of 34 multidrug resistant P. aeruginosa （MRPA）strains for amikacin, ceftazidime, ciprofloxacin, cefoperazone/sulbactam, cefepime, imipenem, levofloxacin, piperacillin/tazobaetam, aztreonam, piperacillin and tobramicin were 14.7 %, 29.4%, 61.8%, 67.6%, 70.6%, 73.5%, 76.5%, 76.5%, 79.4%, 91.2% and 94. 1%, respectively, with 100.0% for others antibiotics. The positive diagnosis rates of gene encoding extended-spectrum β-lactamases FOX, IMP, DHA, and VIM were 23. 5%, 11. 8%, 11. 8% and 5. 9%, 24 （70. 6%） were deleted in outer membrane protein gene oprD2, other β-lactamases genes were absent in all isolates, the type Ⅰ integrating enzyme gene int Ⅰ 1 was positive in 34 strains. CONCLUSIONS P. aeruginosa carries various β-lactamases genes in clinical PAE patients, and the deletion ratio of oprD2 gene is high, type T integrating enzyme exists largely in MRPA.