Center for Biomedical Research, Health Research Organization, National Research and Innovation Agency, Cibinong Science Center-JL. Raya Jakarta-Bogor Km. 46, Bogor 16911, West Jawa Province, Indonesia.
Indonesian Research Center for Veterinary Science, Agricultural Research and Development Agency, Indonesian Ministry of Agriculture, JL. RE. Martadinata 30, Bogor 16114, West Jawa Province, Indonesia.
J Parasitol. 2023 Aug 1;109(4):436-444. doi: 10.1645/21-95.
Identifying a trypanosome isolate is generally based on morphological observations and molecular identification of one of the genes, usually internal transcribed spacer 1 and 2 of ribosomal DNA (ITS1 rDNA, ITS2 rDNA), a variant surface glycoprotein of Rode Trypanozoon antigen type 1.2 (VSG RoTat 1.2), or expression site-associated genes (ESAG). However, this identification is insufficient because these genes cannot distinguish organisms in the subgenus Trypanozoon to the species level. A molecular approach using at least 5 sets of primers is needed, namely, ITS1, ESAG6/7, MINI, RoTat 1.2, and ND5, for stratified selection to obtain more targeted and conclusive results. Using this method to analyze isolates from Indonesia provided unexpected results: 9 isolates previously identified as Trypanozoon were found to have the kDNA maxicircle gene. Nine isolates of Trypanosoma equiperdum were identified for the first time in Indonesia, isolated from bovine (cattle and buffaloes). The identification of T. equiperdum in the 9 isolates was confirmed by analysis of the nucleotide sequence identity of the nad5-kDNA maxicircle gene.
鉴定一种锥虫分离株通常基于形态学观察和对核糖体 DNA（ITS1 rDNA、ITS2 rDNA）的一个基因、 Rode Trypanozoon 抗原型 1.2 的变异表面糖蛋白（VSG RoTat 1.2）或表达位点相关基因（ESAG）的分子鉴定。然而，这种鉴定是不够的，因为这些基因无法将亚属锥虫中的生物区分到种的水平。需要使用至少 5 组引物的分子方法，即 ITS1、ESAG6/7、MINI、RoTat 1.2 和 ND5，进行分层选择，以获得更有针对性和结论性的结果。使用这种方法分析来自印度尼西亚的分离株，得到了意想不到的结果：此前鉴定为锥虫的 9 个分离株被发现具有 kDNA 大环线基因。首次在印度尼西亚发现了 9 株伊氏锥虫，它们从牛（牛和水牛）中分离出来。通过分析 nad5-kDNA 大环线基因的核苷酸序列同一性，确认了这 9 个分离株中存在伊氏锥虫。
