Çelikbilek Nevreste, Gözalan Ayşegül, Özdem Birsen, Kırca Fisun, Açıkgöz Ziya Cibali
Ankara Atatürk Education and Research Hospital, Medical Microbiology Laboratory, Ankara, Turkey.
Mikrobiyol Bul. 2015 Apr;49(2):259-65. doi: 10.5578/mb.9031.
The aim of the study was to evaluate the change of the frequency of extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae isolates from urine samples of outpatients in years and to analyse the antibiotic resistance profiles for a rational drug use. The urine samples cultured in our laboratory from the patients who were admitted to outpatient clinics of our hospital between years 2007-2013 were included in this study. Enterobacteriaceae strains were isolated and identified by conventional methods and API 20E system (BioMérieux, France). The standard antimicrobial susceptibility tests were performed by Kirby Bauer disk diffusion method. ESBL production were screened by double-disk synergy method according to CLSI guidelines. E-test method (BioMérieux, France) were used for the verification of suspicious ESBL production. The identification and antimicrobial susceptibility testing were performed for a total of 12.535 isolates. Of the isolates 8716 were identified as Escherichia coli (69.3%), 1514 were Klebsiella pneumoniae/oxytoca (12.1%), 257 were Proteus mirabilis (2.1%), 345 were other Enterobacteriae members (8%), 411 were various non-fermentative gram-negative bacteria (3.3%) and 1292 were various gram-positive bacteria (10.3%). The total positivity rate of ESBL was found as 21.8% (2.283/10.487), and the ESBL positive rates for E.coli, K.pneumoniae/oxytoca and P.mirabilis were 21.2%, 28.2% and 4.7%, respectively. Other Enterobacteriaceae isolates were not evaluated because of the absence of standardized methods and breakpoint values. There was no statistically significant difference among ESBL producing isolates within seven years (p= 0.364). The antibiotic resistance rates of the ESBL-positive isolates were statistically higher than ESBL-negative isolates [amoxicillin-clavulanate (73.1%/11.3%), trimethoprim-sulfamethoxazole (63.1%/31.0%), nitrofurantoin (17.3%/8.6%), gentamicin (42.2%/10.1%), amikacin (3.5%/0.9%), tobramisin (56.8%/10.5%), imipenem (0.3%/0.1%), ofloxacin (66.8%/19.8%), ticarcillin-clavulanate (73.5%/19.8%), piperacillin-tazobactam (28.8%/5.0%)] (p< 0.05). Statistically significant variations were detected within the years for the resistance rates of amoxicillin-clavulanate (p= 0.001), tobramycin (p=0.003), ofloxacin (p= 0.001), ticarcillin-clavulanate (p= 0.001) and piperacillin-tazobactam (p= 0.001) were detected within the years. Although a quite high percentage of ESBL positivity in Enterobacteriaceae isolates was determined, there was a slight but not statistically significant increase of this value during the seven-year period. The stability of the percentage of ESBL positivity may indicate a positive change in the habit of the usage of beta-lactam antibiotics. According to the results of our study, the most effective drugs for ESBL-producing isolates were piperacillin-tazobactam among inhibitor combinations, amikacin among aminoglycosides and nitrofurantoin among orally-used drugs.
本研究的目的是评估多年来门诊患者尿液样本中产超广谱β-内酰胺酶(ESBL)肠杆菌科分离株的频率变化,并分析抗生素耐药谱以促进合理用药。本研究纳入了2007年至2013年期间在我院门诊就诊患者的尿液样本,这些样本在我们实验室进行培养。采用常规方法和API 20E系统(法国生物梅里埃公司)分离和鉴定肠杆菌科菌株。采用 Kirby Bauer 纸片扩散法进行标准抗菌药物敏感性试验。根据CLSI指南,采用双纸片协同法筛选ESBL的产生。使用E-test法(法国生物梅里埃公司)验证可疑的ESBL产生情况。共对12535株分离株进行了鉴定和抗菌药物敏感性试验。其中8716株鉴定为大肠埃希菌(69.3%),1514株为肺炎克雷伯菌/产酸克雷伯菌(12.1%),257株为奇异变形杆菌(2.1%),345株为其他肠杆菌科成员(8%),411株为各种非发酵革兰阴性菌(3.3%),1292株为各种革兰阳性菌(10.3%)。ESBL的总阳性率为21.8%(2283/10487),大肠埃希菌、肺炎克雷伯菌/产酸克雷伯菌和奇异变形杆菌的ESBL阳性率分别为21.2%、28.2%和4.7%。由于缺乏标准化方法和折点值,未对其他肠杆菌科分离株进行评估。7年内产ESBL的分离株之间无统计学显著差异(p = 0.364)。ESBL阳性分离株的抗生素耐药率在统计学上高于ESBL阴性分离株[阿莫西林-克拉维酸(73.1%/11.3%)、甲氧苄啶-磺胺甲恶唑(63.1%/31.0%)、呋喃妥因(17.3%/8.6%)、庆大霉素(42.2%/10.1%)、阿米卡星(3.5%/0.9%)、妥布霉素(56.8%/10.5%)、亚胺培南(0.3%/0.1%)、氧氟沙星(66.8%/19.8%)、替卡西林-克拉维酸(73.5%/19.8%)、哌拉西林-他唑巴坦(28.8%/5.0%)](p < 0.05)。在各年份中,阿莫西林-克拉维酸(p = 0.001)、妥布霉素(p = 0.003)、氧氟沙星(p = 0.001)、替卡西林-克拉维酸(p = 0.001)和哌拉西林-他唑巴坦(p = 0.001)的耐药率存在统计学显著差异。尽管在肠杆菌科分离株中确定了相当高比例的ESBL阳性,但在7年期间该值有轻微但无统计学显著的增加。ESBL阳性率的稳定性可能表明β-内酰胺类抗生素使用习惯的积极变化。根据我们的研究结果,对于产ESBL的分离株,在抑制剂联合用药中最有效的药物是哌拉西林-他唑巴坦,在氨基糖苷类药物中是阿米卡星,在口服药物中是呋喃妥因。