Schulthess Bettina, Bloemberg Guido V, Zbinden Andrea, Mouttet Forouhar, Zbinden Reinhard, Böttger Erik C, Hombach Michael
Institute of Medical Microbiology, University of Zurich, Zurich, Switzerland
Institute of Medical Microbiology, University of Zurich, Zurich, Switzerland.
J Clin Microbiol. 2016 Mar;54(3):543-8. doi: 10.1128/JCM.03107-15. Epub 2015 Dec 9.
Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has entered clinical laboratories, facilitating identification of bacteria. Here, we evaluated the MALDI Biotyper (Bruker Daltonics) for the identification of fastidious Gram-negative rods (GNR). Three sample preparation methods, direct colony transfer, direct transfer plus on-target formic acid preparation, and ethanol-formic acid extraction, were analyzed for 151 clinical isolates. Direct colony transfer applied with the manufacturer's interpretation criteria resulted in overall species and genus identification rates of 43.0% and 32.5%, respectively; 23.2% of the isolates were not identified, and two misidentifications (1.3%) were observed. The species identification rates increased to 46.4% and 53.7% for direct transfer plus formic acid preparation and ethanol-formic acid extraction, respectively. In addition, we evaluated score value cutoff alterations. The identification rates hardly increased by reducing the genus cutoff, while reducing the 2.0 species cutoff to 1.9 and to 1.8 increased the identification rates to up to 66.2% without increasing the rate of misidentifications. This study shows that fastidious GNR can reliably be identified using the MALDI Biotyper. However, the identification rates do not reach those of nonfastidious GNR such as the Enterobacteriaceae. In addition, two approaches optimizing the identification of fastidious GNR by the MALDI Biotyper were demonstrated: formic acid-based on-target sample treatment and reductions in cutoff scores to increase the species identification rates.
基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)已进入临床实验室,有助于细菌鉴定。在此,我们评估了MALDI Biotyper(布鲁克道尔顿公司)对苛养革兰氏阴性杆菌(GNR)的鉴定能力。对151株临床分离株分析了三种样品制备方法,即直接菌落转移法、直接转移加靶上甲酸制备法和乙醇-甲酸提取法。按照制造商的解释标准采用直接菌落转移法,总体菌种和菌属鉴定率分别为43.0%和32.5%;23.2%的分离株未得到鉴定,观察到两例假阳性鉴定(1.3%)。直接转移加甲酸制备法和乙醇-甲酸提取法的菌种鉴定率分别提高到46.4%和53.7%。此外,我们评估了评分值截断点的改变。降低菌属截断点时鉴定率几乎没有增加,但将菌种截断点从2.0降低到1.9和1.8时,鉴定率提高到了66.2%,且未增加假阳性鉴定率。本研究表明,使用MALDI Biotyper能够可靠地鉴定苛养GNR。然而其鉴定率未达到肠杆菌科等非苛养GNR的鉴定率水平。此外,还展示了两种优化MALDI Biotyper对苛养GNR鉴定的方法:基于甲酸的靶上样品处理和降低截断分数以提高菌种鉴定率。