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A new Trypanosoma cruzi genotyping method enables high resolution evolutionary analyses

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. Sev...

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Autores principales: Probst, Christian Macagnan, Melo, Myllena de Fátima Alheiros Dias, Pavoni, Daniela Parada, Toledo, Max Jean de Ornelas, Galdino, Tainah Silva, Brandão, Adeilton Alves, Britto, Constança, Krieger, Marco Aurelio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Instituto Oswaldo Cruz, Ministério da Saúde 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8405150/
https://www.ncbi.nlm.nih.gov/pubmed/34468503
http://dx.doi.org/10.1590/0074-02760200538
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author Probst, Christian Macagnan
Melo, Myllena de Fátima Alheiros Dias
Pavoni, Daniela Parada
Toledo, Max Jean de Ornelas
Galdino, Tainah Silva
Brandão, Adeilton Alves
Britto, Constança
Krieger, Marco Aurelio
author_facet Probst, Christian Macagnan
Melo, Myllena de Fátima Alheiros Dias
Pavoni, Daniela Parada
Toledo, Max Jean de Ornelas
Galdino, Tainah Silva
Brandão, Adeilton Alves
Britto, Constança
Krieger, Marco Aurelio
author_sort Probst, Christian Macagnan
collection PubMed
description 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.
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spelling pubmed-84051502021-09-09 A new Trypanosoma cruzi genotyping method enables high resolution evolutionary analyses Probst, Christian Macagnan Melo, Myllena de Fátima Alheiros Dias Pavoni, Daniela Parada Toledo, Max Jean de Ornelas Galdino, Tainah Silva Brandão, Adeilton Alves Britto, Constança Krieger, Marco Aurelio Mem Inst Oswaldo Cruz Original Article 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. Instituto Oswaldo Cruz, Ministério da Saúde 2021-08-30 /pmc/articles/PMC8405150/ /pubmed/34468503 http://dx.doi.org/10.1590/0074-02760200538 Text en https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License
spellingShingle Original Article
Probst, Christian Macagnan
Melo, Myllena de Fátima Alheiros Dias
Pavoni, Daniela Parada
Toledo, Max Jean de Ornelas
Galdino, Tainah Silva
Brandão, Adeilton Alves
Britto, Constança
Krieger, Marco Aurelio
A new Trypanosoma cruzi genotyping method enables high resolution evolutionary analyses
title A new Trypanosoma cruzi genotyping method enables high resolution evolutionary analyses
title_full A new Trypanosoma cruzi genotyping method enables high resolution evolutionary analyses
title_fullStr A new Trypanosoma cruzi genotyping method enables high resolution evolutionary analyses
title_full_unstemmed A new Trypanosoma cruzi genotyping method enables high resolution evolutionary analyses
title_short A new Trypanosoma cruzi genotyping method enables high resolution evolutionary analyses
title_sort new trypanosoma cruzi genotyping method enables high resolution evolutionary analyses
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8405150/
https://www.ncbi.nlm.nih.gov/pubmed/34468503
http://dx.doi.org/10.1590/0074-02760200538
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