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肠球菌临床分离株的药敏试验及菌种鉴定

Antimicrobial susceptibility and species identification for clinical isolates of enterococci.

作者信息

Hsieh S R

机构信息

Department of Medical Technology, Chungtai Institute of Health Sciences and Technology, Taichung, Taiwan, ROC.

出版信息

J Microbiol Immunol Infect. 2000 Dec;33(4):253-7.

Abstract

In order to describe the susceptibility patterns and determine the identities of Enterococcus spp. isolated at a regional teaching hospital in Taichung during the period from June through November 1998, 96 clinical isolates of enterococci were collected for further analysis. Major sources of these isolates included urine, wound, and pus. Species identification was performed using the API 20 Strep system and supplemental tests. The minimum inhibitory concentrations (MICs) of six antimicrobial agents were determined by E-test for each isolate. Disk diffusion tests were also performed and the results were compared with those reported by the clinical laboratory. Because gentamicin susceptibility tests showed inconsistent results in many isolates, MIC determinations by the microbroth dilution method were also performed for these isolates. All isolates were tested for beta-lactamase production using the chromogenic method. The results showed that Enterococcus faecalis was the most frequently encountered species (86.5%), followed by Enterococcus faecium (7.3%), Enterococcus avium (5.2%) and Enterococcus casseliflavus (1.0%). The MIC90 of ampicillin, penicillin, vancomycin, teicoplanin, ciprofloxacin and gentamicin for total isolates were 1, 3, 2, 0.25, 1, and more than 1024 microg/mL, respectively. All isolates were susceptible to vancomycin and teicoplanin. The same rate of resistance (3.1%) was found to penicillin, ampicillin and ciprofloxacin in all isolates. There were 50 (52%) and 48 (50%) isolates with high level streptomycin and gentamicin resistance, respectively. The MIC90 of ampicillin and penicillin for E. faecium were significantly higher than those for other species (96 and > 256 vs. < or = 1 and < or = 3 microg/mL, respectively); in fact, ampicillin or penicillin-resistance was only found in E. faecium. No organism was found to produce beta-lactamase. There were 29 isolates showing discrepant results between the study findings and clinical laboratory report for gentamicin susceptibility. Most isolates (27/ 29) reported as susceptible to gentamicin by the clinical laboratory showed a high level gentamicin resistance by MIC determinations. The inconsistent results may be due to discrepancy in interpretations for heterogeneous resistance, bias in selecting colonies for the disk diffusion test, or variation in the quality of Mueller-Hinton agar used. The results suggest that any pure isolates growing some colonies within the inhibition zone should be considered as gentamicin resistant even if the zone diameter is equal or greater than the susceptible breakpoint. In order to obtain accurate results, the gentamicin MIC should be determined by the dilution method for enterococcal isolates that yield intermediate inhibition zones or zones just above the susceptible limit.

摘要

为描述台中某区域教学医院1998年6月至11月期间分离出的肠球菌属的药敏模式并确定其种类,收集了96株临床分离的肠球菌进行进一步分析。这些分离株的主要来源包括尿液、伤口和脓液。使用API 20 Strep系统及补充试验进行菌种鉴定。采用E试验测定每株分离菌对6种抗菌药物的最低抑菌浓度(MIC)。同时进行纸片扩散试验,并将结果与临床实验室报告的结果进行比较。由于许多分离株的庆大霉素药敏试验结果不一致,因此也采用微量肉汤稀释法对这些分离株进行MIC测定。采用显色法对所有分离株进行β-内酰胺酶产生情况检测。结果显示,粪肠球菌是最常见的菌种(86.5%),其次是屎肠球菌(7.3%)、鸟肠球菌(5.2%)和格氏肠球菌(1.0%)。所有分离株对氨苄西林、青霉素、万古霉素、替考拉宁、环丙沙星和庆大霉素的MIC90分别为1、3、2、0.25、1和大于1024μg/mL。所有分离株对万古霉素和替考拉宁敏感。所有分离株对青霉素、氨苄西林和环丙沙星的耐药率相同(3.1%)。分别有50株(52%)和48株(50%)分离株对链霉素和庆大霉素高度耐药。屎肠球菌对氨苄西林和青霉素的MIC90显著高于其他菌种(分别为96和>256μg/mL,而其他菌种≤1和≤3μg/mL);事实上,仅在屎肠球菌中发现对氨苄西林或青霉素耐药。未发现有菌株产生β-内酰胺酶。有29株分离株在庆大霉素药敏试验中研究结果与临床实验室报告存在差异。临床实验室报告为对庆大霉素敏感的大多数分离株(27/29)经MIC测定显示对庆大霉素高度耐药。结果不一致可能是由于对异质性耐药的解释存在差异、纸片扩散试验中菌落选择存在偏差或所用Mueller-Hinton琼脂质量存在差异。结果表明,即使抑菌圈内生长有一些菌落的纯培养分离株的抑菌圈直径等于或大于敏感界值,也应视为对庆大霉素耐药。为获得准确结果,对于产生中度抑菌圈或抑菌圈刚好高于敏感界限的肠球菌分离株,应采用稀释法测定庆大霉素MIC。

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