Ojima-Kato Teruyo, Yamamoto Naomi, Iijima Yoshio, Tamura Hiroto
Knowledge Hub Aichi, Aichi Science and Technology Foundation, Yakusa, Toyota, Aichi 470-0356, Japan.
School of Agriculture, Meijo University, Shiogamaguchi, Tenpaku-ku, Nagoya, Aichi 468-0073, Japan.
J Microbiol Methods. 2015 Dec;119:233-8. doi: 10.1016/j.mimet.2015.11.005. Epub 2015 Nov 10.
O157, O26, and O111 are the most important O serogroups of enterohemorrhagic Escherichia coli worldwide. Recently we reported a strategy for discriminating these serotypes from the others using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) based on the S10-spc-alpha operon gene-encoded ribosomal protein mass spectrum (S10-GERMS) method. To realize the fully automated identification of microorganisms at species- or serotype-level with the concept of S10-GERMS method, novel software named Strain Solution for MALDI-TOF MS was developed. In this study, the Strain Solution was evaluated with a total of 45 E. coli isolates including O26, O91, O103, O111, O115, O121, O128, O145, O157, O159, and untyped serotypes. The Strain Solution could accurately discriminate 92% (11/12) of O157 strains, 100% (13/13) of O26 and O111 strains from the others with three biomarkers in an automated manner. In addition, this software could identify 2 different E. coli strains (K-12 as a non-O157 representative and O157) in mixed samples. The results suggest that Strain Solution will be useful for species- or serotype-level classification of microorganisms in the fields of food safety and diagnostics.
O157、O26和O111是全球范围内肠出血性大肠杆菌最重要的O血清型。最近,我们报道了一种基于S10-spc-α操纵子基因编码的核糖体蛋白质谱(S10-GERMS)方法,利用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)将这些血清型与其他血清型区分开来的策略。为了基于S10-GERMS方法的概念实现微生物在种水平或血清型水平的全自动鉴定,开发了名为MALDI-TOF MS菌株解决方案的新型软件。在本研究中,使用包括O26、O91、O103、O111、O115、O121、O128、O145、O157、O159及未分型血清型在内的总共45株大肠杆菌分离株对该菌株解决方案进行了评估。该菌株解决方案能够以自动化方式通过三种生物标志物准确地将92%(11/12)的O157菌株、100%(13/13)的O26和O111菌株与其他菌株区分开来。此外,该软件能够识别混合样品中的2种不同大肠杆菌菌株(作为非O157代表的K-12和O157)。结果表明,菌株解决方案将有助于食品安全和诊断领域微生物的种水平或血清型水平分类。