Please use this identifier to cite or link to this item:
https://www.arca.fiocruz.br/handle/icict/68227
Type
ArticleCopyright
Open access
Collections
- INI - Artigos de Periódicos [3646]
- IOC - Artigos de Periódicos [12980]
Metadata
Show full item record
UTILITY OF MAGNETIC BEAD-BASED AUTOMATED DNA EXTRACTION TO IMPROVE CHAGAS DISEASE MOLECULAR DIAGNOSIS
Author
Affilliation
The University of Texas at El Paso. Department of Biological Sciences. El Paso, TX, USA.
The University of Texas at El Paso. Department of Biological Sciences. El Paso, TX, USA.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia e Parasitologia Molecular. Plataforma de Análise Molecular. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.
The University of Texas at El Paso. Department of Biological Sciences. El Paso, TX, USA.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia e Parasitologia Molecular. Plataforma de Análise Molecular. Rio de Janeiro, RJ, Brasil.
The University of Texas at El Paso. Department of Biological Sciences. El Paso, TX, USA.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia e Parasitologia Molecular. Plataforma de Análise Molecular. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Rio de Janeiro, RJ, Brasil.
The University of Texas at El Paso. Department of Biological Sciences. El Paso, TX, USA.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Virologia e Parasitologia Molecular. Plataforma de Análise Molecular. Rio de Janeiro, RJ, Brasil.
Abstract
Chagas disease, caused by Trypanosoma cruzi, remains a significant global health challenge, particularly in the molecular diagnostics of low parasitemia during the chronic phase. This highlights the critical need for enhanced diagnostic methodologies. In response, this study evaluates the effectiveness of an automated magnetic beads-based DNA extraction method in improving the molecular diagnosis of Chagas disease compared to the traditional silica column-based extraction. Accordingly, this research seeks to enhance the DNA yield, purity, and sensitivity of real-time PCR (qPCR) assays for detecting T. cruzi satDNA. Blood samples spiked with guanidine–EDTA solution and varying concentrations of T. cruzi were used to compare the two extraction methods. The results indicated that the magnetic bead-based method outperformed the silica column in terms of DNA concentration, purity, and earlier detection of T. cruzi satDNA. Although both methods had similar limits of detection at a 95% confidence interval, the magnetic bead-based approach demonstrated higher sensitivity and reproducibility, particularly in low-parasitemia samples. The findings suggest that the magnetic beads-based DNA extraction method offers a more reliable, faster, and more sensitive alternative for diagnosing chronic Chagas disease, potentially improving clinical outcomes by enabling more accurate and earlier parasite detection.
Share