Research of antigenic diversity and phylogenetic marker genes for Babesia microti

本研究在日本啮齿动物中新鉴定或建立了几种准otsu - ssurdna型（长野ssurdna型）和us - ssurdna型巴贝虫分离株或菌株，以及kobe - ssurdna型和otsu - ssurdna型巴贝虫。长野- ssurdna型微型贝氏菌仅在安氏大鳍鱼中鉴定到，而在长野田鼠种中鉴定到otsu - ssurdna型微型贝氏菌，准otsu - ssurdna型和otsu - ssurdna型贝氏菌是当地特有的。我们还从台湾啮齿类动物中鉴定或建立了kobe - ssurdna型microti（梅山分离株）的分离株或菌株。对这些分离株和已建立的Gl、N/A、Kobe、Otsu和Hachimaki菌株进行了ITS1/ITS2序列的研究。不同ssurdna类型间ITS1/ITS2的同源性约为50-60%。在otsu - ssurdna型中，各菌株的ITS1和ITS2序列仅相差1 ~ 3个核苷酸。对于微小贝氏杆菌的kobe - ssurdna型，所有日本鼠分离物均具有完全相同的ITS1/1TS2序列。神户菌株与眉山菌株的同源性为92%左右。对于us - ssurdna型微杆菌，GI和N/A菌株ITS序列完全相同，而日本分离株ITS1/ITS2序列与GI和N/A菌株有明显的差异，同源性为91.1% ~ 97.2%。经免疫印迹分析，神户菌株和大津菌株的主要免疫反应抗原分别约为42kD和35kD。不同ITS1/ITS2序列的同一ssurdna型微恙螨主要免疫反应抗原的分子量似乎有所不同，但间接荧光抗体检测显示不同ITS1/ITS2序列的同一ssurdna型微恙螨具有交叉免疫反应性。由于无法克隆主要的免疫反应性抗原，无法对分离株或菌株的免疫反应性抗原进行更详细的结构分析。少

In this study, we newly identified or established several isolates or strains of quasi-Otsu-SSUrDNA-type (Nagano SSUrDNA-type) and US-SSUrDNA-type Babesia microti in addition to Kobe-SSUrDNA-type and Otsu-SSUrDNA-type of B. microti from Japanese rodents. Nagano-SSUrDNA-type B. microti was exclusively identified in Eothenomys andersoni, while Otsu-SSUrDNA-type B. microti was identified in Apodemus species in Nagano, where quasi-Otsu-and Otsu-SSUrDNA-type B. microti are endemic. We also identified or established isolates or strains of Kobe-SSUrDNA-type B. microti (the Meishan isolate) from Taiwanese rodents.For these isolates or strains plus the Gl, the N/A, the Kobe, the Otsu and the Hachimaki strains, which had been previously established, the sequences of ITS1/ITS2 were investigated. The percent identities of ITS1/ITS2 between the different SSUrDNA-types were around 50-60%. Within the Otsu-SSUrDNA-type, the ITS1 and ITS2 sequences of all B. microti isolates were different only by 1-3 … More nucleotides from each other. For the Kobe-SSUrDNA-type of B. microti, all of the isolates from Japanese rodents possessed completely identical ITS1/1TS2 sequences. However, between the Kobe and Meishan strains, around 92% identities were showed. For the US-SSUrDNA-type of B.microti, the ITS sequences of the GI and N/A strains were completely identical, while the ITS1/ITS2 sequence of the Japanese isolate was clearly differentiated from those of the GI and N/A strains with the 91.1 %-97.2% identities.The main immunoreactive antigens of the Kobe and Otsu strains were examined by immunoblot analysis and were shown to be approximately 42kD and 35kD, respectively. Between the same SSUrDNA-type B. microti with different ITS1/ITS2 sequences, the molecular weights of main immunoreactive antigens seemed to be somewhat different, although indirect fluorescent antibody test showed cross immunoreactivities between the same SSUrDNA-type B. microti with different ITS1/ITS2 sequences. Because we could not clone the main immunoreactive antigens, more detailed structural analysis for immunoreactive antigens of the isolates or strains was impossible. Less