Detection of Trypanosoma lewisi from wild rats in Southern China and its genetic diversity based on the ITS1 and ITS2 sequences

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• 作者：Hai-Jun Tang^a ; You-Gen Lan^a ; Yan-Zi Wen^a ; Xi-Chen Zhang^b ; Marc Desquesnes^c ; ^d ; Ting-Bao Yang^a ; Geoff Hide^e ; Zhao-Rong Lun^a ; ^e ; ^{lsslzr@mail.sysu.edu.cn}
• 关键词：Trypanosoma lewisi ; Rattus norvegicus ; PCR ; Loop-mediated isothermal amplification (LAMP) ; ITS ; Phylogenetic tree
• 刊名：Infection, Genetics and Evolution
• 出版年：2012
• 出版时间：July, 2012
• 年：2012
• 卷：12
• 期：5
• 页码：1046-1051
• 全文大小：305 K

文摘

Trypanosoma lewisi has widely been considered as a non-pathogenic rat trypanosome. However, more and more cases of humans infected with T. lewisi have been reported around the world, indicating that it can infect humans in some undetermined circumstances. Quick and sensitive diagnosis of infection by T. lewisi is important for both treatment of patients and epidemiological studies of this parasite. In this paper, three methods i.e. wet blood smear (diagnosis by microscopy), PCR and LAMP were used to detect T. lewisi from 238 wild rats (Rattus norvegicus) collected from the field in Huadu, Guangdong province, China. Infection rates of these samples detected by the 3 methods was 6.7 % (16/238), 12.6 % (30/238), and 18.9 % (45/238), respectively. LAMP could detect all samples shown positive by microscopical observation of wet smear and by single PCR indicating good potential for application in the detection of T. lewisi. So far as we know, this is the first report of the LAMP method being used to detect T. lewisi in wild rats. The specific T. lewisi LAMP primers were able to amplify the target fragment from the genomic DNA of 19 T. lewisi strains isolated from Huadu, Guangdong province (n = 16), Changchun, Jilin province of China (n = 1) and from Thailand (n = 2). Based on the analyses of ITS1 (internal transcribed spacer 1) and ITS2 sequences, these 19 strains show a very close genetic relationship with over 96-97 % similarity to the other corresponding sequences of T. lewisi published in Genbank. Phylogenetic trees of the species in the subgenus Herpetosoma were constructed, based on the ITS1 and ITS2 sequences, and these results also indicate that they are closely related and in the same clade.