目前登革热的诊断方法,如血清学测试,在与其他病毒的交叉反应性方面具有局限性。为了解决这个问题,我们探索了将环介导等温扩增(LAMP)技术与适体亲和力相结合开发即时检测的潜力.在这项研究中,我们使用了60份血清样本。能够与登革热病毒结合的适体被用作捕获遗传物质的平台。并将其性能与商业套件进行了比较。通过RT-PCR和比色逆转录环介导等温扩增(RT-LAMP)检测登革病毒,允许在不需要设备的情况下目视观察结果。在适体LAMP测定的背景下,我们的分析显示,在60个样本中,有38个检测到登革热病毒,与RT-PCR和/或APTA-RT-PCR相比,具有95%的灵敏度和100%的特异性。重要的是,我们在评估寨卡病毒阳性样本时没有观察到交叉反应,强调测定的选择性。这种与RT-LAMP(APTA-RT-LAMP)方法结合的病毒RNA的创新适体捕获具有以更简单和更快的方式为被忽视的传染病提供有价值的分子见解的潜力。
目的:登革热是一种被忽视的热带病,在热带和亚热带国家具有重要的流行病学意义。目前对这种感染的诊断提出了挑战,如血清学试验中的交叉反应性。寻找方法来加强这种疾病的诊断是至关重要的,鉴于缺乏特定的治疗方法。一个准确的,简单,有效的诊断有助于改善感染者的管理。在这种情况下,我们的工作结合了分子生物学技术,如等温回路扩增,用适体检测生物样本中的登革热病毒。我们的方法根据颜色变化产生比色结果,结果在不到2小时内可用。此外,与分子PCR测试相比,它需要更简单的设备。
Current diagnostic methods for dengue, such as serological tests, have limitations in terms of cross-reactivity with other viruses. To address this issue, we explored the potential of combining the loop-mediated isothermal amplification (LAMP) technique with the affinity of aptamers to develop point-of-care testing. In this study, we utilized 60 serum samples. An aptamer capable of binding to the dengue virus was employed as a platform for capturing genetic material, and its performance was compared to a commercial kit. Dengue virus was detected through RT-PCR and colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP), allowing visual observation of the results without the need for equipment. In the context of the aptamer LAMP assay, our analysis revealed the detection of the dengue virus in 38 out of 60 samples, with 95% sensitivity and 100% specificity compared to RT-PCR and/or APTA-RT-PCR. Importantly, we observed no cross-reaction when assessing samples positive for the zika virus, underscoring the assay\'s selectivity. This innovative aptameric capture of the viral RNA in combination with the RT-LAMP (APTA-RT-LAMP) method has the potential to offer valuable molecular insights into neglected infectious diseases in a simpler and faster manner.
OBJECTIVE: Dengue is a neglected tropical disease of significant epidemiological importance in tropical and subtropical countries. Current diagnostics for this infection present challenges, such as cross-reactivity in serological tests. Finding ways to enhance the diagnosis of this disease is crucial, given the absence of specific treatments. An accurate, simple, and effective diagnosis contributes to the improved management of infected individuals. In this context, our work combines molecular biology techniques, such as isothermal loop amplification, with aptamers to detect the dengue virus in biological samples. Our method produces colorimetric results based on a color change, with outcomes available in less than 2 hours. Moreover, it requires simpler equipment compared to molecular PCR tests.