Ashizawa Kazuho, Murata Syota, Terada Takashi, Ito Daisuke, Bunya Masaru, Watanabe Koji, Teruuchi Yoko, Tsuchida Sachio, Satoh Mamoru, Nishimura Motoi, Matsushita Kazuyuki, Sugama Yuji, Nomura Fumio
R&D Department, Nittobo Medical Co., Ltd., 1 Shiojima, Fukuhara, Fukuyama, Koriyama, Fukushima 963-8061, Japan.
Division of Laboratory Medicine, Clinical Genetics and Proteomics, Chiba University Hospital, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan; Division of Clinical Mass Spectrometry, Chiba University Hospital, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan.
J Microbiol Methods. 2017 Aug;139:54-60. doi: 10.1016/j.mimet.2017.04.013. Epub 2017 Apr 29.
Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) can be used to identify pathogens in blood culture samples. However, sample pretreatment is needed for direct identification of microbes in blood culture bottles. Conventional protocols are complex and time-consuming. Therefore, in this study, we developed a method for collecting bacteria using polyallylamine-polystyrene copolymer for application in wastewater treatment technology. Using representative bacterial species Escherichia coli and Staphylococcus capitis, we found that polyallylamine-polystyrene can form visible aggregates with bacteria, which can be identified using MALDI-TOF MS. The processing time of our protocol was as short as 15min. Hemoglobin interference in MALDI spectra analysis was significantly decreased in our method compared with the conventional method. In a preliminary experiment, we evaluated the use of our protocol to identify clinical isolates from blood culture bottles. MALDI-TOF MS-based identification of 17 strains from five bacterial species (E. coli, Klebsiella pneumoniae, Enterococcus faecalis, S. aureus, and S. capitis) collected by our protocol was satisfactory. Prospective large-scale studies are needed to further evaluate the clinical application of this novel and simple method of collecting bacteria in blood culture bottles.
基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)可用于鉴定血培养样本中的病原体。然而,要直接鉴定血培养瓶中的微生物,需要进行样本预处理。传统方法复杂且耗时。因此,在本研究中,我们开发了一种使用聚烯丙胺-聚苯乙烯共聚物收集细菌的方法,用于废水处理技术。使用代表性细菌物种大肠杆菌和头状葡萄球菌,我们发现聚烯丙胺-聚苯乙烯能与细菌形成可见聚集体,可通过MALDI-TOF MS进行鉴定。我们方法的处理时间短至15分钟。与传统方法相比,我们的方法在MALDI光谱分析中血红蛋白干扰显著降低。在初步实验中,我们评估了使用我们的方法鉴定血培养瓶中的临床分离株。通过我们的方法收集的来自五种细菌物种(大肠杆菌、肺炎克雷伯菌、粪肠球菌、金黄色葡萄球菌和头状葡萄球菌)的17株菌株,基于MALDI-TOF MS的鉴定结果令人满意。需要进行前瞻性大规模研究,以进一步评估这种在血培养瓶中收集细菌的新颖且简单方法的临床应用。