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Title: PCR-based identification of Trypanosoma lewisi and Trypanosoma musculi using maxicircle kinetoplast DNA
Authors: Hong, Xiao-Kun 
Zhang, Xuan 
Fusco, Octavio Alejandro 
Lan, You-Gen 
Lun, Zhao-Rong 
Lai, De-Hua 
Keywords: Maxicircle kinetoplast DNA;Trypanosoma lewis;;Trypanosoma musculi;Morfología
Issue Date: Jul-2017
Journal: Acta tropica 
Trypanosoma lewisi, transmitted by rat fleas, is a widespread pathogen specific to rats with records of human infection cases. Its closely related species with global distribution, Trypanosoma musculi, is transmitted between mice by ingestion of infected fleas. These trypanosomes are of similar morphology, making it difficult to distinguish them by microscopy. In this study, we have developed a rapid, sensitive and reliable PCR method for the diagnosis of T. lewisi and T. musculi. The T. lewisi-specific amplicons were not produced by other Trypanosoma, such as T. musculi, T. brucei complex or T. cruzi, neither by an outgroup of Leishmania amazonensis. The detection limits of the three pairs of T. lewisi-specific primers were 50ng, 1ng and 10ng of total DNA, respectively. The primers designed for T. musculi primers showed specifically that amplicon strictly in T. musculi and their detection limits were 10ng and 1ng of total DNA. To simplify the detection process, we managed to apply our method directly on tail blood samples without complicated DNA purification. In conclusion, PCR with our primers could be a highly sensitive, specific protocol to detect and distinguish T. lewisi and T. musculi from other trypanosomes.
DOI: 10.1016/j.actatropica.2017.04.007
Appears in Collections:Publicaciones INP

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