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首页> 外文期刊>Frontiers in Microbiology >Applicability of an in-House Saponin-Based Extraction Method in Bruker Biotyper Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry System for Identification of Bacterial and Fungal Species in Positively Flagged Blood Cultures
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Applicability of an in-House Saponin-Based Extraction Method in Bruker Biotyper Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry System for Identification of Bacterial and Fungal Species in Positively Flagged Blood Cultures

机译：内部基于皂苷的提取方法在Bruker Biotyper基质辅助的激光解吸/电离飞行时间质谱系统中用于鉴定正标记血液培养物中细菌和真菌种类的适用性

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We used an in-house saponin-based extraction method to evaluate the performance of the Bruker Biotyper matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) system for the identification of bacteria and fungi in 405 positively flagged blood culture bottles. Results obtained from MALDI-TOF/MS were compared with those obtained using conventional phenotypic identification methods. Of the 405 positively flagged blood culture bottles, 365 showed monomicrobal growth and were correctly identified to the species (72.1%) or genus (89.6%) level using the Bruker Biotyper system. The remaining 40 positively flagged blood culture bottles showed polymicrobial growth. Of them, 82.5% ( n = 33) of the isolates were correctly identified to the species level and 92.5% ( n = 37) to the genus level using the Bruker Biotyper system. The overall accuracy of identification to the genus level in flagged blood cultures was 89.5% for Gram-positive organisms, 93.5% for Gram-negative pathogens and 71.9% for fungi. Confidence scores were ≥1.500 for 307 (75.8%) bottles, ≥1.700 for 249 (61.5%) bottles and ≥2.000 for 142 (35.1%) bottles. None of the yeast cultures yielded scores ≥1.700. Using an identification-score cutoff of ≥1.500, the MALDI Biotyper correctly identified 99.2% of Gram-positive bacteria, 97.6% of Gram-negative bacteria and 100% of yeast isolates to the genus level and 77.6% of Gram-positive bacteria, 87.1% of Gram-negative bacteria and 100.0% of yeast isolates to the species level. The overall rate of identification using our protocol was 89.9% (364/405) for genus level identification and 73.1% (296/405) for species level identification. Yeast isolates yielded the lowest confidence scores, which compromised the accuracy of identification. Further optimization of the protein extraction procedure in positive blood cultures is needed to improve the rate of identification.
机译：我们使用了一种基于内部皂苷的提取方法来评估Bruker Biotyper基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF / MS）系统的性能，以鉴定405中的细菌和真菌正面标记的血液培养瓶。将从MALDI-TOF / MS获得的结果与使用常规表型鉴定方法获得的结果进行比较。在405个带有阳性标记的血液培养瓶中，有365个显示出微生物的生长，并使用Bruker Biotyper系统正确鉴定了该物种（72.1％）或属（89.6％）的水平。其余40个带有正向标记的血液培养瓶显示出微生物的生长。其中，使用Bruker Biotyper系统可以正确鉴定出82.5％（n = 33）的分离物至物种水平，正确鉴定出92.5％（n = 37）的属水平。在标记的血液培养物中，鉴定到属水平的总体准确性对于革兰氏阳性生物是89.5％，对于革兰氏阴性病原体是93.5％，对于真菌是71.9％。置信分数为307（75.8％）瓶≥1.500、249（61.5％）瓶≥1.700和142（35.1％）瓶≥2.000。没有一个酵母培养物得分≥1.700。使用≥1.500的鉴定分数截止值，MALDI Biotyper正确鉴定出99.2％的革兰氏阳性菌，97.6％的革兰氏阴性菌和100％的酵母菌属属水平和77.6％的革兰氏阳性菌87.1 ％的革兰氏阴性菌和100.0％的酵母菌分离到物种水平。使用我们的方案进行鉴定的总体比率，属水平鉴定为89.9％（364/405），而物种鉴定为73.1％（296/405）。酵母分离株产生最低的置信度得分，这损害了鉴定的准确性。需要在阳性血液培养物中进一步优化蛋白质提取程序，以提高鉴定率。

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