Standardization of DNA extraction from paraffinized spleen samples: molecular diagnosis of human malaria
Abstract Background Plasmodium vivax is the main species responsible for human malaria in Brazil, and one of its manifestations is splenic malaria, though there are still challenges in its diagnosis. The present study aimed to standardize Plasmodium sp. DNA extraction from histological slices of spl...
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ftdoajarticles:oai:doaj.org/article:6d994beffb34423c92510009070812df 2024-01-07T09:41:49+01:00 Standardization of DNA extraction from paraffinized spleen samples: molecular diagnosis of human malaria Raimunda Sandra Pacheco Souza Monique F. dos Reis Luiz Carlos de Lima Ferreira Manuela C. Morais Antonio Kassio S. Lima Laila Rowena Albuquerque Barbosa Gisely Cardoso de Melo Marcus Vinicius Guimaraes de Lacerda 2023-11-01T00:00:00Z https://doi.org/10.1186/s12936-023-04764-3 https://doaj.org/article/6d994beffb34423c92510009070812df EN eng BMC https://doi.org/10.1186/s12936-023-04764-3 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-023-04764-3 1475-2875 https://doaj.org/article/6d994beffb34423c92510009070812df Malaria Journal, Vol 22, Iss 1, Pp 1-6 (2023) Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2023 ftdoajarticles https://doi.org/10.1186/s12936-023-04764-3 2023-12-10T01:48:34Z Abstract Background Plasmodium vivax is the main species responsible for human malaria in Brazil, and one of its manifestations is splenic malaria, though there are still challenges in its diagnosis. The present study aimed to standardize Plasmodium sp. DNA extraction from histological slices of spleen and diagnosis using real-time qPCR. Methods This study performed a microtomy of a paraffin-embedded spleen as a positive control for P. vivax from a patient who had been previously diagnosed with the parasite. The sample was deparaffinized with xylol and ethanol, then DNA extraction was performed with two commercial kits. qPCR was carried out with the Taqman system for detection of Plasmodium sp. and was made species-specific using PvmtCOX1 gene. From 2015 to 2019, 200 spleen samples were obtained from trauma patients subjected to splenectomy in Manaus, Amazonas. All the samples were tested for cell-free human DNA (cfDNA). Results The deparaffinization and the Plasmodium vivax DNA extraction method was successfully standardized, and the control sample was positive for P. vivax. Of the 200 samples, all qPCRs were negative, but they were positive for human PCR. Conclusion Paraffinization is practical and efficient for the preservation of samples, but the formation of bonds between proteins and DNA makes extraction difficult. Despite this, in this study, it was possible to standardize a method of DNA extraction for detecting P. vivax. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 22 1 |
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Directory of Open Access Journals: DOAJ Articles |
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English |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Raimunda Sandra Pacheco Souza Monique F. dos Reis Luiz Carlos de Lima Ferreira Manuela C. Morais Antonio Kassio S. Lima Laila Rowena Albuquerque Barbosa Gisely Cardoso de Melo Marcus Vinicius Guimaraes de Lacerda Standardization of DNA extraction from paraffinized spleen samples: molecular diagnosis of human malaria |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Plasmodium vivax is the main species responsible for human malaria in Brazil, and one of its manifestations is splenic malaria, though there are still challenges in its diagnosis. The present study aimed to standardize Plasmodium sp. DNA extraction from histological slices of spleen and diagnosis using real-time qPCR. Methods This study performed a microtomy of a paraffin-embedded spleen as a positive control for P. vivax from a patient who had been previously diagnosed with the parasite. The sample was deparaffinized with xylol and ethanol, then DNA extraction was performed with two commercial kits. qPCR was carried out with the Taqman system for detection of Plasmodium sp. and was made species-specific using PvmtCOX1 gene. From 2015 to 2019, 200 spleen samples were obtained from trauma patients subjected to splenectomy in Manaus, Amazonas. All the samples were tested for cell-free human DNA (cfDNA). Results The deparaffinization and the Plasmodium vivax DNA extraction method was successfully standardized, and the control sample was positive for P. vivax. Of the 200 samples, all qPCRs were negative, but they were positive for human PCR. Conclusion Paraffinization is practical and efficient for the preservation of samples, but the formation of bonds between proteins and DNA makes extraction difficult. Despite this, in this study, it was possible to standardize a method of DNA extraction for detecting P. vivax. |
format |
Article in Journal/Newspaper |
author |
Raimunda Sandra Pacheco Souza Monique F. dos Reis Luiz Carlos de Lima Ferreira Manuela C. Morais Antonio Kassio S. Lima Laila Rowena Albuquerque Barbosa Gisely Cardoso de Melo Marcus Vinicius Guimaraes de Lacerda |
author_facet |
Raimunda Sandra Pacheco Souza Monique F. dos Reis Luiz Carlos de Lima Ferreira Manuela C. Morais Antonio Kassio S. Lima Laila Rowena Albuquerque Barbosa Gisely Cardoso de Melo Marcus Vinicius Guimaraes de Lacerda |
author_sort |
Raimunda Sandra Pacheco Souza |
title |
Standardization of DNA extraction from paraffinized spleen samples: molecular diagnosis of human malaria |
title_short |
Standardization of DNA extraction from paraffinized spleen samples: molecular diagnosis of human malaria |
title_full |
Standardization of DNA extraction from paraffinized spleen samples: molecular diagnosis of human malaria |
title_fullStr |
Standardization of DNA extraction from paraffinized spleen samples: molecular diagnosis of human malaria |
title_full_unstemmed |
Standardization of DNA extraction from paraffinized spleen samples: molecular diagnosis of human malaria |
title_sort |
standardization of dna extraction from paraffinized spleen samples: molecular diagnosis of human malaria |
publisher |
BMC |
publishDate |
2023 |
url |
https://doi.org/10.1186/s12936-023-04764-3 https://doaj.org/article/6d994beffb34423c92510009070812df |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 22, Iss 1, Pp 1-6 (2023) |
op_relation |
https://doi.org/10.1186/s12936-023-04764-3 https://doaj.org/toc/1475-2875 doi:10.1186/s12936-023-04764-3 1475-2875 https://doaj.org/article/6d994beffb34423c92510009070812df |
op_doi |
https://doi.org/10.1186/s12936-023-04764-3 |
container_title |
Malaria Journal |
container_volume |
22 |
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1 |
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1787422615041736704 |