Research of antigenic diversity and phylogenetic marker genes for Babesia microti

在本研究中，除了日本啮齿动物的Kobe-SSUrDNA型和Otsu-SSUrDNA型外，我们还新鉴定或建立了几个准Otsu-SSUrDNA型(长野SSUrDNA型)和US-SSUrDNA型巴贝斯虫。Nagano-SSUrDNA型B.microti仅在Eothenomys andersoni中被发现，而Otsu-SSUrDNA型B.microti在长野姬鼠中被发现，其中准Otsu和Otsu-SSUrDNA型B.microti是长野特有的。我们还鉴定或建立了台湾啮齿动物的Kobe-SSUrDNA型B.microti分离株(眉山分离株)，并对这些分离株和已建立的G1、N/A、Kobe、Otsu和Hachimaki株进行了ITS1/ITS2的序列分析。不同SSUrDNA类型间ITS1/ITS2的同源性在50-60%左右。在OTSU-SSUrDNA型中，所有微小芽孢杆菌的ITS1和ITS2序列仅有1-3…的差异彼此之间有更多的核苷酸。对于Kobe-SSUrDNA型B.microti，所有来自日本啮齿动物的分离株都具有完全相同的ITS1/1TS2序列。然而，神户株和眉山株之间显示出约92%的同源性。对于US-SSUrDNA型B.microti，GI株和N/A株的ITS序列完全相同，而日本株的ITS1/ITS2序列与GI株和N/A株的ITS1/ITS2序列有91.1%-97.2%的同源性。免疫印迹分析表明，神户株和大津株的主要免疫活性抗原分别约为42kD和35kD。具有不同ITS1/ITS2序列的相同SSUrDNA型B.microti之间，主要免疫反应抗原的分子量似乎略有不同，尽管间接荧光抗体试验显示相同SSUrDNA型B.microti与不同ITS1/ITS2序列之间存在交叉免疫反应。由于我们不能克隆主要的免疫反应抗原，所以不可能对分离株或菌株的免疫反应抗原进行更详细的结构分析。较少

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