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Applications of copolymer for rapid identification of bacteria in blood culture broths using matrix-assisted laser desorption ionization time-of-flight mass spectrometry

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
Journal of microbiological methods 2017 v.139 pp. 54-60
Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Staphylococcus capitis, bacteria, blood, bottles, composite polymers, culture media, hemoglobin, matrix-assisted laser desorption-ionization mass spectrometry, wastewater treatment
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.