Aktaş Zerrin, Gönüllü Nevriye, Schneider Ines, Bal Ciğdem, Bauernfeind Adolf
Istanbul Universitesi Istanbul Tip Fakültesi, Mikrobiyoloji ve Klinik Mikrobiyoloji Anabilim Dali, Istanbul.
Mikrobiyol Bul. 2005 Oct;39(4):421-9.
In this study, a CTX-M type extended spectrum beta-lactamase (ESBL) enzyme has been detected in a multiresistant Escherichia coli strain which was isolated from the urine sample of a hospitalized patient. Minimum inhibitor concentrations (MIC) of the tested antibiotics were determined by the agar dilution technique according to the Clinical and Laboratory Standards Institute (CLSI, formerly NCCLS) guidelines. The isolate was found to be sensitive to imipinem, moderately susceptible to chloramphenicol and resistant to ceftazidime, cefotaxime, aztreonam, ciprofloxacin, tobramycin, tetracycline and trimethoprim/sulphamethoxazole. Ceftazidime/ceftazidime-clavulanic acid and cefotaxime/cefotaxime-clavulanic acid rates were found as >8 and the results were accepted as positive for an ESBL. The MIC of cefotaxime (256 microg/ml) was found four fold higher than that of ceftazidime (64 microg/ml) and the production of a CTX-M- type ESBL was investigated in the strain. Cefotaxime resistance, together with tobramycin and tetracycline resistance, was transferred to the recipient strain by conjugation. The pl's of the culture extracts of the isolate were found as 5.4, 7.5 and 9.1 by isoelectric focusing (IEF) method, but cefotaxime was hydrolysed only by the beta-lactamase focusing at a pl of 9.1 in the following bioassay. The bla-gene was amplified with the CTX-M group specific primers and sequencing of the polymerase chain reaction (PCR) product proved the enzyme to be CTX-M-15. This isolate was also found to harbor TEM- and SHV-type and OXA-10-like ESBLs, by IEF and PCR.
在本研究中,从一名住院患者的尿液样本中分离出的一株多重耐药大肠杆菌菌株中检测到了CTX-M型超广谱β-内酰胺酶(ESBL)。根据临床和实验室标准协会(CLSI,原NCCLS)指南,采用琼脂稀释技术测定了受试抗生素的最低抑菌浓度(MIC)。该分离株对亚胺培南敏感,对氯霉素中度敏感,对头孢他啶、头孢噻肟、氨曲南、环丙沙星、妥布霉素、四环素和甲氧苄啶/磺胺甲恶唑耐药。头孢他啶/头孢他啶-克拉维酸和头孢噻肟/头孢噻肟-克拉维酸比率>8,结果被判定为ESBL阳性。头孢噻肟的MIC(256μg/ml)比头孢他啶(64μg/ml)高4倍,对该菌株进行了CTX-M型ESBL产生情况的研究。通过接合试验,头孢噻肟耐药性以及妥布霉素和四环素耐药性被转移至受体菌株。通过等电聚焦(IEF)法测定,该分离株培养提取物的等电点分别为5.4、7.5和9.1,但在随后的生物测定中,仅等电点为9.1的β-内酰胺酶能水解头孢噻肟。用CTX-M组特异性引物扩增bla基因,聚合酶链反应(PCR)产物测序证明该酶为CTX-M-15。通过IEF和PCR还发现该分离株携带TEM型、SHV型和OXA-10样ESBL。