Farani Priscila S G, Lopez Jacqueline, Faier-Pereira Amanda, Hasslocher-Moreno Alejandro Marcel, Almeida Igor C, Moreira Otacilio C
Department of Biological Sciences, The University of Texas at El Paso, El Paso, TX 79902, USA.
Platform of Molecular Analysis, Laboratory of Molecular Virology and Parasitology, Oswaldo Cruz Institute, Oswaldo Cruz Foundation, Rio de Janeiro 21040-360, RJ, Brazil.
Int J Mol Sci. 2025 Jan 23;26(3):937. doi: 10.3390/ijms26030937.
Chagas disease, caused by , 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 satDNA. Blood samples spiked with guanidine-EDTA solution and varying concentrations of 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 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.
由[病原体名称未给出]引起的恰加斯病仍然是一项重大的全球卫生挑战,尤其是在慢性期低寄生虫血症的分子诊断方面。这凸显了对改进诊断方法的迫切需求。作为回应,本研究评估了一种基于自动磁珠的DNA提取方法与传统基于硅胶柱的提取方法相比,在改善恰加斯病分子诊断方面的有效性。因此,本研究旨在提高用于检测[病原体名称未给出]卫星DNA的实时PCR(qPCR)检测的DNA产量、纯度和灵敏度。用添加了胍-EDTA溶液和不同浓度[病原体名称未给出]的血样来比较这两种提取方法。结果表明,基于磁珠的方法在DNA浓度、纯度以及对[病原体名称未给出]卫星DNA的更早检测方面优于硅胶柱法。尽管两种方法在95%置信区间的检测限相似,但基于磁珠的方法表现出更高的灵敏度和可重复性,尤其是在低寄生虫血症样本中。研究结果表明,基于磁珠的DNA提取方法为诊断慢性恰加斯病提供了一种更可靠、更快且更灵敏的替代方法,有可能通过实现更准确和更早的寄生虫检测来改善临床结果。