Meng Xinan, Ou Yonglin, Jiang Wen, Guo Yaqiong, Xiao Lihua, Feng Yaoyu, Li Na
State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, South China Agricultural University, Guangzhou 510642, China.
State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, South China Agricultural University, Guangzhou 510642, China - Guangdong Laboratory for Lingnan Modern Agriculture, Center for Emerging and Zoonotic Diseases, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
Parasite. 2025;32:6. doi: 10.1051/parasite/2025002. Epub 2025 Jan 31.
In addition to the ribosomal internal transcribed spacer (ITS) locus, four loci (MS1, MS3, MS4, and MS7) have been identified to develop multilocus sequence typing tools for high-resolution genotyping of Enterocytozoon bieneusi in previous studies. However, the use of only five loci was insufficient for population genetic analysis of E. bieneusi from diverse hosts. In this study, comparison of a clinical genome sequence (C44566) with the whole genome sequence of an E. bieneusi isolate (H348) in GenBank led to the selection of the hypothetical protein 1 (hp1) and tubulin 1 (tub1) loci. Further analysis of the two loci with 156 E. bieneusi-positive samples showed high sequence polymorphisms in ITS Groups 1-6 and 10. Altogether, 30 and 23 sequence types were identified at hp1 and tub1, respectively. Genotyping based on the two loci confirmed the lack of genetic differentiation between Group 1 and Group 2 genotypes, as previously reported. Moreover, the genotypes in Groups 4 and 5 are more divergent from other genotypes within Groups 1-10. However, isolates in Group 11 and 12 could not be amplified at the hp1 and tub1 loci, supporting the previous conclusion of genetic uniqueness of the two genotype groups. The identified genetic markers and generated data could be used to develop a multilocus sequence typing tool for high-resolution genotyping of E. bieneusi, which would also have implications for understanding the taxonomy of Enterocytozoon spp., the public health significance of E. bieneusi in animals, and sources of E. bieneusi infections in humans.
除核糖体内部转录间隔区（ITS）位点外，先前的研究已鉴定出四个位点（MS1、MS3、MS4和MS7），用于开发多位点序列分型工具，以对微小隐孢子虫进行高分辨率基因分型。然而，仅使用五个位点不足以对来自不同宿主的微小隐孢子虫进行群体遗传分析。在本研究中，将临床基因组序列（C44566）与GenBank中微小隐孢子虫分离株（H348）的全基因组序列进行比较，从而选择了假设蛋白1（hp1）和微管蛋白1（tub1）位点。对这两个位点与156份微小隐孢子虫阳性样本的进一步分析显示，ITS第1 - 6组和第10组存在高度的序列多态性。总共在hp1和tub1位点分别鉴定出30种和23种序列类型。基于这两个位点的基因分型证实了第1组和第2组基因型之间缺乏遗传分化，正如先前报道的那样。此外，第4组和第5组的基因型与第1 - 10组内的其他基因型差异更大。然而，第11组和第12组的分离株在hp1和tub1位点无法扩增，支持了先前关于这两个基因型组遗传独特性的结论。所鉴定的遗传标记和生成的数据可用于开发一种多位点序列分型工具，用于微小隐孢子虫的高分辨率基因分型，这也将有助于理解隐孢子虫属的分类学、微小隐孢子虫在动物中的公共卫生意义以及人类微小隐孢子虫感染的来源。
