A capacitive laser-induced graphene based aptasensor for SARS-CoV-2 detection in human saliva.
A capacitive laser-induced graphene based aptasensor for SARS-CoV-2 detection in human saliva.
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DOI：
10.1371/journal.pone.0290256
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发表时间：
2023
期刊：
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
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作者：
SARS-CoV-2病毒引起的COVID-19大流行加速了诊断工具的发展。与电化学生物传感器集成的设备可能是一种有趣的替代方案，以应对对测试的高需求，特别是在缺乏标准检测技术的情况下。作为识别元件的适体由于其稳定性、特异性和对结合靶分子的敏感性而有用。我们已经开发了一种非侵入性的电化学适配子为基础的生物传感器靶向SARS-CoV-2在人类唾液中。该适体有望用于检测SARS-CoV-2野生型及其变异株的Spike蛋白。用生物素标记的适体对涂覆有铂纳米颗粒的激光诱导石墨烯（LIG）电极进行生物功能化。用于BA.1感测的电化学阻抗谱（EIS）在补充有汇集唾液的氯化钠/碳酸氢钠溶液中进行。为了评估传感性能，用10至10，000拷贝/ml的UV减毒病毒体的人工样品测试适体传感器。通过将适体传感器暴露于非靶向病毒（hCoV-OC 43、甲型流感病毒和RSV-A）来评估选择性。进一步使用EIS数据输出来选择合适的响应变量和截止频率。电容响应于衰减的BA的逐渐加载而增加。适体传感器在较低病毒载量（10-100拷贝/ml）下对BA.1具有灵敏度和特异性，并且能够区分阴性和阳性人工样本（对其他病毒具有毒株特异性：OC 43、甲型流感病毒和RSV-A）。适体传感器在人工样本中检测到SARS-CoV-2，估计LOD为1790拷贝/ml。在人类临床样本中，适体传感器的准确性为72%，阳性一致率为75%，阴性一致率为67%。我们的研究结果表明，适体传感器是一种有希望的候选者，可以在病毒体浓度较低的感染早期检测SARS-CoV-2，这可能有助于预防COVID-19的无症状传播。
SARS-CoV-2 virus induced CoVID-19 pandemic has accelerated the development of diagnostic tools. Devices integrated with electrochemical biosensors may be an interesting alternative to respond to the high demand for testing, particularly in contexts where access to standard detection technologies is lacking. Aptamers as recognition elements are useful due to their stability, specificity, and sensitivity to binding target molecules. We have developed a non-invasive electrochemical aptamer-based biosensor targeting SARS-CoV-2 in human saliva. The aptamer is expected to detect the Spike protein of SARS-CoV-2 wildtype and its variants. Laser-induced graphene (LIG) electrodes coated with platinum nanoparticles were biofunctionalized with a biotin-tagged aptamer. Electrochemical Impedance Spectroscopy (EIS) for BA.1 sensing was conducted in sodium chloride/sodium bicarbonate solution supplemented with pooled saliva. To estimate sensing performance, the aptasensor was tested with contrived samples of UV-attenuated virions from 10 to 10,000 copies/ml. Selectivity was assessed by exposing the aptasensor to non-targeted viruses (hCoV-OC43, Influenza A, and RSV-A). EIS data outputs were further used to select a suitable response variable and cutoff frequency. Capacitance increases in response to the gradual loading of the attenuated BA.1. The aptasensor was sensitive and specific for BA.1 at a lower viral load (10–100 copies/ml) and was capable of discriminating between negative and positive contrived samples (with strain specificity against other viruses: OC43, Influenza A, and RSV-A). The aptasensor detected SARS-CoV-2 with an estimated LOD of 1790 copies/ml in contrived samples. In human clinical samples, the aptasensor presents an accuracy of 72%, with 75% of positive percent of agreement and 67% of negative percent of agreement. Our results show that the aptasensor is a promising candidate to detect SARS-CoV-2 during early stages of infection when virion concentrations are low, which may be useful for preventing the asymptomatic spread of CoVID-19.
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