A new Trypanosoma cruzi genotyping method enables high resolution evolutionary analyses

Memorias do Instituto Oswaldo Cruz

article

Christian Macagnan Probst¹  Myllena de Fátima Alheiros Dias Melo²  Daniela Parada Pavoni¹  Max Jean de Ornelas Toledo³  Tainah Silva Galdino⁴  Adeilton Alves Brandão⁴  Constança Britto²  Marco Aurelio Krieger¹ 

[1] Instituto Carlos Chagas, Laboratório de Genômica Funcional;Instituto Oswaldo Cruz, Laboratório de Biologia Molecular e Doenças Endêmicas;Universidade Estadual de Maringá, Departamento de Ciências da Saúde, Laboratório de Doença de Chagas;Instituto Oswaldo Cruz, Laboratório Interdisciplinar de Pesquisas Médicas

关键词: Trypanosoma cruzi;    genotyping techniques;    high-throughput nucleotide sequencing;    trans-sialidase;   

DOI  :  10.1590/0074-02760200538

学科分类：地质学

来源: Bioline International

【 摘 要 】

BACKGROUND Trypanosoma cruzi is an important human pathogen in Latin America with nearly seven million people infected. It has a large degree of genetic diversity, classified into six discrete typing units (DTUs), which probably influences its physiological behavior and clinical manifestations. Several genotyping methods are available, with distinct performance on easiness, cost, resolution and applicability; no method excels in all parameters. OBJECTIVES AND METHODS To devise a molecular method for T. cruzi genotyping, based on polymerase chain reaction (PCR) amplification of a single target with multiple copies in the nuclear genome by large scale sequencing. We have applied this method to 29 T. cruzi isolates, comprising all described DTUs. FINDINGS We were able to classify all samples into sub DTU level with high robustness. Evolutionary relationship between DTUs were ascertained, suggesting that TcIII and TcIV DTUs are non-hybrid, and DTU IV is more similar to the common ancestral. CONCLUSION As the TS-LSS method is based on a single PCR reaction, comprising several copies of the target, it is probably useful for clinical samples, when the amount of DNA is a limiting factor. As large scale sequencing systems become more common, the TS-LSS method can be increasingly applied for T. cruzi genotyping.

【 授权许可】

Free