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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Instituto Oswaldo Cruz, Ministério da Saúde
2021
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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. |
format | Online Article Text |
id | pubmed-8405150 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Instituto Oswaldo Cruz, Ministério da Saúde |
record_format | MEDLINE/PubMed |
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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