Hamamoto Kouta, Ueda Shuhei, Yamamoto Yoshimasa, Hirai Itaru
Laboratory of Microbiology, School of Health Sciences, Faculty of Medicine, University of the Ryukyus, Okinawa, Japan.
Laboratory of Microbiology, School of Health Sciences, Faculty of Medicine, University of the Ryukyus, Okinawa, Japan Japan Science and Technology Agency/Japan International Cooperation Agency, Science and Technology Research Partnership for Sustainable Development (JST/JICA, SATREPS), Osaka, Japan.
J Clin Microbiol. 2015 Jun;53(6):1848-53. doi: 10.1128/JCM.00227-15. Epub 2015 Mar 25.
Genotyping and characterization of bacterial isolates are essential steps in the identification and control of antibiotic-resistant bacterial infections. Recently, one novel genotyping method using three genomic guided Escherichia coli markers (GIG-EM), dinG, tonB, and dipeptide permease (DPP), was reported. Because GIG-EM has not been fully evaluated using clinical isolates, we assessed this typing method with 72 E. coli collection of reference (ECOR) environmental E. coli reference strains and 63 E. coli isolates of various genetic backgrounds. In this study, we designated 768 bp of dinG, 745 bp of tonB, and 655 bp of DPP target sequences for use in the typing method. Concatenations of the processed marker sequences were used to draw GIG-EM phylogenetic trees. E. coli isolates with identical sequence types as identified by the conventional multilocus sequence typing (MLST) method were localized to the same branch of the GIG-EM phylogenetic tree. Sixteen clinical E. coli isolates were utilized as test isolates without prior characterization by conventional MLST and phylogenetic grouping before GIG-EM typing. Of these, 14 clinical isolates were assigned to a branch including only isolates of a pandemic clone, E. coli B2-ST131-O25b, and these results were confirmed by conventional typing methods. Our results suggested that the GIG-EM typing method and its application to phylogenetic trees might be useful tools for the molecular characterization and determination of the genetic relationships among E. coli isolates.
细菌分离株的基因分型和特性鉴定是识别和控制抗生素耐药性细菌感染的关键步骤。最近,有报道称一种使用三种基因组导向的大肠杆菌标记物(GIG-EM)——dinG、tonB和二肽通透酶(DPP)的新型基因分型方法。由于尚未使用临床分离株对GIG-EM进行全面评估,我们用72株大肠杆菌参考(ECOR)环境大肠杆菌参考菌株和63株不同遗传背景的大肠杆菌分离株对该分型方法进行了评估。在本研究中,我们指定了dinG的768 bp、tonB的745 bp和DPP靶序列的655 bp用于分型方法。处理后的标记序列串联起来用于绘制GIG-EM系统发育树。通过传统多位点序列分型(MLST)方法鉴定出的具有相同序列类型的大肠杆菌分离株位于GIG-EM系统发育树的同一分支上。16株临床大肠杆菌分离株在进行GIG-EM分型之前未通过传统MLST和系统发育分组进行预先鉴定,用作测试分离株。其中,14株临床分离株被分配到一个分支,该分支仅包含大流行克隆大肠杆菌B2-ST131-O25b的分离株,这些结果通过传统分型方法得到了证实。我们的结果表明,GIG-EM分型方法及其在系统发育树上的应用可能是用于大肠杆菌分离株分子特性鉴定和遗传关系确定的有用工具。