Challhua Ronaldo, Akashi Larissa, Zuñiga Jose, Beatriz de Carvalho Ruthner Batista Helena, Moratelli Ricardo, Champi Ana
Center of Natural and Human Sciences, Federal University of ABC, Santo André, SP, 09210-580, Brazil(1).
Pasteur Institute, São Paulo, Brazil.
Biosens Bioelectron. 2023 Jul 15;232:115291. doi: 10.1016/j.bios.2023.115291. Epub 2023 Mar 31.
Rabies is a lethal zoonotic disease caused by rabies virus (RABV) that affects human health and the economy. RABV is transmitted mainly by bats in Brazil, and surveillance in remote areas is hampered by the difficulty of properly collecting samples during fieldwork and the diagnosis is performed in laboratory conditions. Here, we report a portable electrochemical biosensor based on nucleic acid interactions for RABV detection in nasopharyngeal swab samples. The working electrode of the biosensor is composed of reduced graphene oxide (rGO) thin-film immobilized with cDNA through pi-pi stacking to enhance virus detection and specificity. Sensor performance was determined using RNA, and swab samples from bats. RNA detection shows good selectivity, and quantification presents a highly linear calibration curve (R = 0.990) using a concentration range of 0.145-25.39 ng/μL. A LOD of 0.104 ng/μL was reached with a sensitivity of 0.321 μA (ng/μL). RABV detection in nasopharyngeal swab samples showed a good difference of positive sample from negative with a response time in seconds, ultra-fast detection compared to known techniques. Three biosensor groups were identified and named after physic-chemical surface characterization as: GO-1, GO-2, and rGO; with best performance for rGO group due to its sp hybridized network. Thus, we have successfully fabricated a promising electrochemical biosensor for fast in-situ detection of the RABV in swab samples, which can be expanded to other enveloped viruses that have RNA.
狂犬病是一种由狂犬病病毒(RABV)引起的致命人畜共患病,会影响人类健康和经济。在巴西,RABV主要通过蝙蝠传播,偏远地区的监测工作受到实地采样困难的阻碍,且诊断是在实验室条件下进行的。在此,我们报告一种基于核酸相互作用的便携式电化学生物传感器,用于检测鼻咽拭子样本中的RABV。该生物传感器的工作电极由通过π-π堆积固定有cDNA的还原氧化石墨烯(rGO)薄膜组成,以增强病毒检测和特异性。使用RNA和蝙蝠的拭子样本测定了传感器性能。RNA检测显示出良好的选择性,在0.145 - 25.39 ng/μL的浓度范围内进行定量时呈现出高度线性的校准曲线(R = 0.990)。检测限达到0.104 ng/μL,灵敏度为0.321 μA/(ng/μL)。鼻咽拭子样本中的RABV检测显示阳性样本与阴性样本有良好差异,响应时间以秒计,与已知技术相比检测速度超快。通过物理化学表面表征鉴定出三个生物传感器组,并分别命名为:GO - 1、GO - 2和rGO;其中rGO组性能最佳,因其具有sp杂化网络。因此,我们成功制备了一种有前景的电化学生物传感器,用于快速原位检测拭子样本中的RABV,该传感器可扩展用于检测其他具有RNA的包膜病毒。