Perše Gabrijela, Samošćanec Ivana, Bošnjak Zrinka, Budimir Ana, Kuliš Tomislav, Mareković Ivana
Department of Clinical and Molecular Microbiology, University Hospital Centre Zagreb, 10 000 Zagreb, Croatia.
Department of Clinical Microbiology and Hospital Infections, University Hospital "Sveti Duh", 10 000 Zagreb, Croatia.
Life (Basel). 2022 Oct 30;12(11):1744. doi: 10.3390/life12111744.
In order to further accelerate pathogen identification from positive blood cultures (BC), various sample preparation protocols to identify bacteria with MALDI-TOF MS directly from positive BCs have been developed. We evaluated an in-house method in comparison to the Sepsityper® Kit (Bruker Daltonics, Bremen, Germany) as well as the benefit of an on-plate formic acid extraction step following positive signal by the BACTECTM FX system. Confirmation of identification was achieved using subcultured growing biomass used for MALDI-TOF MS analysis. A total of 113 monomicrobial positive BCs were analyzed. The rates of Gram-positive bacteria correctly identified to the genus level using in-house method and Sepsityper® Kit were 63.3% (38/60) and 81.7% (49/60), respectively (p = 0.025). Identification rates at species level for Gram-positive bacteria with in-house method and Sepsityper® kit were 30.0% (18/60) and 66.7% (40/60), respectively (p < 0.001). Identification rates of Gram-negative bacteria were similar with the in-house method and Sepsityper® Kit. Additional on-plate formic acid extraction demonstrated significant improvement in the identification rate of Gram-positive bacteria at both genus and species level for both in-house (p = 0.001, p < 0.001) and Sepsityper® Kit methods (p = 0.007, p < 0.001). Our in-house method is a candidate for laboratory routines with Sepsityper® Kit as a back-up solution when identification of Gram-positive bacteria is unsuccessful.
为了进一步加快从阳性血培养物(BC)中鉴定病原体的速度,人们已经开发出了各种直接从阳性血培养物中用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)鉴定细菌的样本制备方案。我们评估了一种内部方法,并与Sepsityper®试剂盒(德国不来梅布鲁克道尔顿公司)进行了比较,同时还评估了在BACTECTM FX系统出现阳性信号后进行板上甲酸提取步骤的益处。通过用于MALDI-TOF MS分析的传代培养生长生物量来实现鉴定确认。总共分析了113份单一微生物阳性血培养物。使用内部方法和Sepsityper®试剂盒正确鉴定到属水平的革兰氏阳性菌比例分别为63.3%(38/60)和81.7%(49/60)(p = 0.025)。使用内部方法和Sepsityper®试剂盒在种水平上鉴定革兰氏阳性菌的比例分别为30.0%(18/60)和66.7%(40/60)(p < 0.001)。内部方法和Sepsityper®试剂盒对革兰氏阴性菌的鉴定率相似。额外的板上甲酸提取显示,对于内部方法(p = 0.001,p < 0.001)和Sepsityper®试剂盒方法(p = 0.007,p < 0.001),革兰氏阳性菌在属和种水平上的鉴定率都有显著提高。我们的内部方法可作为实验室常规方法的候选方案,当革兰氏阳性菌鉴定不成功时,Sepsityper®试剂盒可作为备用解决方案。