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[铜绿假单胞菌菌株的抗生素敏感性率及β-内酰胺耐药机制]

[Antibiotic susceptibility rates and beta-lactam resistance mechanisms of Pseudomonas aeruginosa strains].

作者信息

Aktaş Zerrin, Satana Dilek, Kayacan Ciğdem, Can Barış, Gönüllü Nevriye, Küçükbasmacı Omer

机构信息

İstanbul University Faculty of İstanbul Medicine, Department of Medical Microbiology, İstanbul, Turkey.

出版信息

Mikrobiyol Bul. 2012 Jul;46(3):386-97.

Abstract

Pseudomonas aeruginosa is a well-known cause of severe and potentially life-threatening infections including bacteremia, skin and wound infections, pulmonary disease, especially among individuals with cycstic fibrosis, nosocomial urinary tract infections, endocarditis and meningitis. The mechanism of resistance to broad-spectrum beta-lactams in P.aeruginosa are overexpression of cephalosporinases and/or class A, B and D beta-lactamases. Recently PER-1 type beta-lactamase has been reported from Turkey, France, Italy, Romania, Hungary, Belgium, Russia, South Korea and India. OXA beta-lactamases have increasingly been reported in clinical strains of P.aeruginosa from various geographical origins. This study was aimed to investigate the antibiotic susceptibility of various P.aeruginosa clinical strains and to define the beta-lactamase enzymes leading to resistance. In this study, a total of 100 P.aeruginosa strains isolated from various clinical specimens (37 urine, 21 blood, 10 sputum, 5 bronchoalveolar lavage, 5 abscess, 5 wound swabs, 4 endotracheal aspirate, 3 throat swabs, 2 catheter tips, one of each pleural and peritoneal fluid) were included. According to Clinical and Laboratory Standards Institute (CLSI) recommendations, susceptibilities of isolates to various antibiotics were investigated by disk diffusion and agar dilution method, and beta-lactamase enzymes were detected by isoelectric focusing (IEF) method. PSE, PER-1, OXA-10-like beta-lactamase genes and MEX-R genes of isolates were investigated by polymerase chain reaction (PCR). According to MIC90 values, the most effective antibiotics were found to be imipenem (8 µg/ml). The MIC90 values of amikacin, ciprofloxacin, cefepime, cefpirome, piperacillin + tazobactam, piperacillin, ceftazidime, ticarcilin, aztreonam and ticarcilin + clavulanic acid were 32, 64, 64, 64, 128/4, 512, 512, 512, 512 and 512/2 µg/ml, respectively. Seven of the isolates were found to be ESBL positive by double-disk synergy method. It was detected that 10% of the isolates were imipenem-susceptible and 9% were intermediate susceptible. Phenotypical investigation of metallo-beta-lactamase enzyme in these strains by MBL E-test method did not reveal a positive result. PER-1 and OXA-10 like beta-lactamases were detected each in 11% of the isolates, and co-presence of PER-like and OXA-10 like enzymes were shown in 4% of the isolates. PSE gene was not found in any of the strains. The MEXR gene was identified in 52% of the isolates. Antibiotic resistance mechanisms in P.aeruginosa strains seems to be complex. Determination of the resistance mechanisms and antibiotic susceptibility rates in P.aeruginosa will guide the proper antimicrobial therapy, reducing the emergence of resistant strains.

摘要

铜绿假单胞菌是严重且可能危及生命的感染的常见病因,包括菌血症、皮肤和伤口感染、肺部疾病,尤其是在囊性纤维化患者中、医院获得性尿路感染、心内膜炎和脑膜炎。铜绿假单胞菌对广谱β-内酰胺类抗生素的耐药机制是头孢菌素酶和/或A、B、D类β-内酰胺酶的过度表达。最近,在土耳其、法国、意大利、罗马尼亚、匈牙利、比利时、俄罗斯、韩国和印度均有PER-1型β-内酰胺酶的报道。在来自不同地理区域的铜绿假单胞菌临床菌株中,OXAβ-内酰胺酶的报道日益增多。本研究旨在调查各种铜绿假单胞菌临床菌株的抗生素敏感性,并确定导致耐药的β-内酰胺酶。在本研究中,共纳入了从各种临床标本中分离出的100株铜绿假单胞菌菌株(37份尿液、21份血液、10份痰液、5份支气管肺泡灌洗液、5份脓肿、5份伤口拭子、4份气管内吸出物、3份咽拭子、2份导管尖端、各1份胸腔积液和腹腔积液)。根据临床和实验室标准协会(CLSI)的建议,采用纸片扩散法和琼脂稀释法研究分离株对各种抗生素的敏感性,采用等电聚焦(IEF)法检测β-内酰胺酶。通过聚合酶链反应(PCR)研究分离株的PSE、PER-1、OXA-10样β-内酰胺酶基因和MEX-R基因。根据MIC90值,发现最有效的抗生素是亚胺培南(8μg/ml)。阿米卡星、环丙沙星、头孢吡肟、头孢匹罗、哌拉西林+他唑巴坦、哌拉西林、头孢他啶、替卡西林、氨曲南和替卡西林+克拉维酸的MIC90值分别为32、64、64、64、128/4、512、512、512、512和512/2μg/ml。通过双纸片协同法发现7株分离株为ESBL阳性。检测发现10%的分离株对亚胺培南敏感,9%为中介敏感。通过MBL E试验法对这些菌株中的金属β-内酰胺酶进行表型研究未发现阳性结果。11%的分离株中分别检测到PER-1和OXA-10样β-内酰胺酶,4%的分离株中同时存在PER样和OXA-10样酶。在任何菌株中均未发现PSE基因。52%的分离株中鉴定出MEXR基因。铜绿假单胞菌菌株的抗生素耐药机制似乎很复杂。确定铜绿假单胞菌的耐药机制和抗生素敏感率将指导合理的抗菌治疗,减少耐药菌株的出现。

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