Yoshida Haruno, Kim Jae-Seok, Maeda Takahiro, Goto Mieko, Tsuyuki Yuzo, Shizuno Kenichi, Takahashi Takashi
Laboratory of Infectious Diseases, Graduate School of Infection Control Sciences and Ōmura Satoshi Memorial Institute, Kitasato University, Tokyo, Japan.
Department of Laboratory Medicine, Kangdong Sacred Heart Hospital, Hallym University College of Medicine, Seoul, Korea.
Ann Lab Med. 2025 May 1;45(3):300-311. doi: 10.3343/alm.2024.0501. Epub 2025 Mar 20.
Multilocus sequence typing (MLST) is well-established for but remains undeveloped for . We established MLST for using isolates from humans and companion animals in Japan and Korea to gain insights into its population biology.
We analyzed 39 and 22 isolates from companion animals and humans, respectively. We selected seven housekeeping genes-, and -used in MLST. Primer pairs for PCR amplification and sequencing were designed based on conserved sites in 10 whole-genome sequences. We determined fragment sequences, variable sites, allelic profiles, and sequence types (STs) of each isolate. A phylogenetic tree of concatenated sequences was constructed using the goeBURST algorithm to identify STs and clonal complexes (CCs). , encoding outer membrane protein A, was genotyped for molecular characterization.
The sequenced fragment lengths and allele numbers of the seven genes were 424, 451, 483, 439, 429, 419, and 440 bp and 16, 13, 15, 18, 22, 19, and 18, respectively. ST1-ST47, including CC2, CC10, CC18, CC31, and CC33, were diversely distributed among the isolates from different hosts/countries. In the seven-gene phylogenetic tree, apart from , all isolates clustered together. goeBURST diagrams revealed diverse ST distributions among different hosts (animal/human) and countries (Japan/Korea/ others). We found clusters 1-4 in genotyping, indicating that MLST discrimination is higher than typing discrimination.
We established MLST for isolates from humans and companion animals in Japan and Korea, thereby providing a robust tool for population biology studies.
多位点序列分型（MLST）在[具体对象1]方面已得到充分确立，但在[具体对象2]方面仍未得到充分发展。我们利用日本和韩国人类及伴侣动物的分离株建立了针对[具体对象2]的MLST，以深入了解其群体生物学特性。
我们分别分析了来自伴侣动物和人类的39株和22株分离株。我们选择了七个管家基因——[基因名称1]、[基因名称2]和[基因名称3]——用于[具体对象2]的MLST。基于10个全基因组序列中的保守位点设计了用于PCR扩增和测序的引物对。我们确定了每个分离株的片段序列、可变位点、等位基因谱和序列类型（STs）。使用goeBURST算法构建串联序列的系统发育树，以识别STs和克隆复合体（CCs）。对编码外膜蛋白A的[基因名称4]进行基因分型以进行分子特征分析。
七个基因的测序片段长度和等位基因数量分别为424、451、483、439、429、419和440 bp以及16、13、15、18、22、19和18个。ST1 - ST47，包括CC2、CC10、CC18、CC31和CC33，在来自不同宿主/国家的分离株中分布多样。在七基因系统发育树中，除了[基因名称5]外，所有分离株聚在一起。goeBURST图揭示了不同宿主（动物/人类）和国家（日本/韩国/其他）之间不同的ST分布。我们在[基因名称4]基因分型中发现了1 - 4簇，表明MLST的鉴别能力高于[基因名称4]分型的鉴别能力。
我们为日本和韩国人类及伴侣动物的[具体对象2]分离株建立了MLST，从而为群体生物学研究提供了一个强大的工具。
