Gastelbondo-Pastrana Bertha, Flórez Luis, Guzmán Camilo, Torres Karina, Garay Evelin, Ballesteros-Villamizar Jolaime, Gutierrez Rosa, la Hoz Daniel Echeverri-De, López Yésica, Contreras Héctor, Arrieta Germán, Serrano-Coll Héctor, Martínez Caty, Pájaro-Castro Nérlis, Arroyo-Salgado Bárbara, Rivero-Herrera Ricardo, Hurtado Eliana, Araújo João Pessoa, Mattar Salim
Instituto de Investigaciones Biológicas del Trópico, Universidad de Córdoba, Campus Berastegui, Córdoba, Colombia; Grupo de Investigaciones Microbiológicas y Biomédicas de Córdoba-GIMBIC, Universidad de Córdoba, Montería, Colombia; Grupo de Salud Pública y Auditoría en Salud, Corporación Universitaria del Caribe-CECAR, Sincelejo, Colombia.
Instituto de Investigaciones Biológicas del Trópico, Universidad de Córdoba, Campus Berastegui, Córdoba, Colombia.
J Virol Methods. 2025 May;334:115116. doi: 10.1016/j.jviromet.2025.115116. Epub 2025 Feb 14.
During the COVID-19 pandemic, reagents for SARS-CoV-2 detection were scarce or sold at high prices, particularly in Latin America. In this study, a significant step towards self-sufficiency was achieved through the development of an in-house extraction kit for detecting SARS-CoV-2 from nasopharyngeal swab samples. The purity and concentration of the RNA extracted using the in-house kit were compared to those obtained using the GeneJET RNA Purification Kit (Thermo-Scientific®) as a reference. The applicability of the RNA extracted using the kit was evaluated using four samples positive for SARS-CoV-2 by NGS sequencing with Illumina®. There were no significant differences between the results obtained with the in-house kit and those obtained with the commercial kit. These findings confirm that the in-house protocol demonstrated satisfactory diagnostic accuracy for detecting the virus in patients with COVID-19. The in-house extraction kit works effectively, providing optimal RNA extraction for genomic characterization and lineage assignment of SARS-CoV-2 within the four positive samples analyzed. This phenol-free kit represents a local design and production achievement, offering an effective solution for RNA extraction and detection and sequencing of SARS-CoV-2 from nasopharyngeal swabs. The data highlight the essential contribution of this study to health and biotechnological sovereignty in Colombia.
在新冠疫情期间，用于检测严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的试剂稀缺或价格高昂，在拉丁美洲尤其如此。在本研究中，通过开发一种用于从鼻咽拭子样本中检测SARS-CoV-2的内部提取试剂盒，在实现自给自足方面迈出了重要一步。将使用内部试剂盒提取的RNA的纯度和浓度与使用GeneJET RNA纯化试剂盒（赛默飞世尔科技公司®）作为对照所获得的结果进行比较。使用Illumina®通过NGS测序对四个SARS-CoV-2阳性样本评估该试剂盒提取的RNA的适用性。内部试剂盒获得的结果与商业试剂盒获得的结果之间没有显著差异。这些发现证实，内部方案在检测新冠患者体内病毒方面显示出令人满意的诊断准确性。内部提取试剂盒工作有效，在所分析的四个阳性样本中为SARS-CoV-2的基因组特征分析和谱系分型提供了最佳的RNA提取。这种无酚试剂盒代表了一项本地设计和生产成果，为从鼻咽拭子中提取、检测和测序SARS-CoV-2提供了有效的解决方案。这些数据突出了本研究对哥伦比亚健康和生物技术主权的重要贡献。
