Laboratory of Microbiology, Ghent University, K.L. Ledeganckstraat 35, B-9000 Ghent, Belgium.
Antimicrob Agents Chemother. 2010 Jun;54(6):2567-74. doi: 10.1128/AAC.00407-10. Epub 2010 Apr 12.
In a small-scale harmonization study involving nine laboratories in eight European countries, the intra- and interlaboratory performances of two commercially available systems, i.e., the VetMIC microplate system and Etest, for antimicrobial susceptibility testing of nonenterococcal lactic acid bacteria (NELAB) and bifidobacteria were analyzed. In addition, one laboratory also performed standard broth microdilution as a reference method. MICs of tetracycline, erythromycin, ampicillin, gentamicin, clindamycin, and streptomycin for the type strains of 25 species of NELAB and bifidobacteria and MICs of vancomycin for a selection of relevant taxa were determined. The previously described lactic acid bacterium susceptibility test medium (LSM) and related mixed-medium formulations, all including Iso-Sensitest broth as a basic component, were used as test media. The overall agreement of median MIC ranges +/- 1 log(2) dilution determined by the VetMIC and Etest methods with the median MICs determined by the reference method was very good for tetracycline, ampicillin, and streptomycin (92.3 to 100%) but low for erythromycin (19.5 to 30.7%) and clindamycin (50.0 to 80.8%). There was a consensus among the participating laboratories that VetMIC was preferred over Etest because of its lower cost, better growth support, and more uniform criteria for MIC end point reading. With the range for acceptable intralaboratory reproducibility being defined as the median MIC +/- 1 log(2) dilution, VetMIC results (with 69.2% of all data sets in the acceptable range) were shown to display greater reproducibility than Etest results (with 58.8% of all data sets in the acceptable range). Also at the interlaboratory level, the proportion of MIC values obtained with VetMIC that belonged to the complete agreement category (60.0%) was higher than the proportion of such values obtained with Etest (47.0%), which indicates a higher degree of interlaboratory reproducibility for the former method. Apart from some agent-specific effects, the majority of VetMIC and Etest replicate data sets were situated within a 1- to 2-log(2) dilution range, suggesting that the two methods can be considered to be equivalent for recognizing resistance phenotypes. This multicenter study has further validated the standard use of LSM and related mixed-medium formulations with commercially available systems and formed the basis for the ongoing development of the ISO 10932/IDF 223 standard for susceptibility testing of NELAB and bifidobacteria.
在一项涉及欧洲八国九家实验室的小规模协调研究中,分析了两种市售系统,即 VetMIC 微孔板系统和 Etest,对非肠球菌乳酸杆菌(NELAB)和双歧杆菌的抗菌药敏试验的实验室内和实验室间性能。此外,一个实验室还进行了标准肉汤微量稀释作为参考方法。测定了 25 种 NELAB 和双歧杆菌的标准株的四环素、红霉素、氨苄西林、庆大霉素、克林霉素和链霉素的 MIC 值,以及相关分类单元的万古霉素 MIC 值。先前描述的乳酸细菌药敏试验培养基(LSM)和相关的混合培养基配方,均包含 Iso-Sensitest 肉汤作为基本成分,用作试验培养基。 VetMIC 和 Etest 方法测定的中位数 MIC 范围 +/- 1 log(2)稀释与参考方法测定的中位数 MIC 非常吻合,对于四环素、氨苄西林和链霉素(92.3%至 100%),而红霉素(19.5%至 30.7%)和克林霉素(50.0%至 80.8%)则较低。参与实验室达成共识,认为 VetMIC 优于 Etest,因为 VetMIC 成本更低、生长支持更好、MIC 终点读取标准更统一。将可接受的实验室内可重复性范围定义为中位数 MIC +/- 1 log(2)稀释, VetMIC 结果(所有数据集中有 69.2%在可接受范围内)显示出比 Etest 结果(所有数据集中有 58.8%在可接受范围内)更高的重现性。在实验室间水平, VetMIC 获得的 MIC 值属于完全一致类别的比例(60.0%)高于 Etest(47.0%),这表明前者方法的实验室间重现性更高。除了一些特定于试剂的影响外, VetMIC 和 Etest 重复数据集中的大多数都位于 1 到 2 log(2)稀释范围内,这表明这两种方法可用于识别耐药表型。这项多中心研究进一步验证了使用市售系统对 LSM 和相关混合培养基配方的标准应用,并为正在进行的 NELAB 和双歧杆菌药敏试验的 ISO 10932/IDF 223 标准的开发奠定了基础。