Chiu Chan-Wen, Xian Minghan, Stephany Jenna L, Xia Xinyi, Chiang Chao-Ching, Ren Fan, Tsai Cheng-Tse, Shan Siang-Sin, Liao Yu-Te, Esquivel-Upshaw Josephine F, Rananaware Santosh R, Nguyen Long T, Macaluso Nicolas C, Jain Piyush K, Cash Melanie N, Mavian Carla N, Salemi Marco, Leon Marino E, Chang Chin-Wei, Lin Jenshan, Pearton Stephen J
Department of Chemical Engineering, University of Florida, Gainesville, Florida 32611.
Department of Restorative Dental Sciences, University of Florida, Gainesville, Florida 32610.
J Vac Sci Technol B Nanotechnol Microelectron. 2022 Mar;40(2):023204. doi: 10.1116/6.0001615. Epub 2022 Feb 9.
The SARS-CoV-2 pandemic has had a significant impact worldwide. Currently, the most common detection methods for the virus are polymerase chain reaction (PCR) and lateral flow tests. PCR takes more than an hour to obtain the results and lateral flow tests have difficulty with detecting the virus at low concentrations. In this study, 60 clinical human saliva samples, which included 30 positive and 30 negative samples confirmed with RT-PCR, were screened for COVID-19 using disposable glucose biosensor strips and a reusable printed circuit board. The disposable strips were gold plated and functionalized to immobilize antibodies on the gold film. After functionalization, the strips were connected to the gate electrode of a metal-oxide-semiconductor field-effect transistor on the printed circuit board to amplify the test signals. A synchronous double-pulsed bias voltage was applied to the drain of the transistor and strips. The resulting change in drain waveforms was converted to digital readings. The RT-PCR-confirmed saliva samples were tested again using quantitative PCR (RT-qPCR) to determine cycling threshold (Ct) values. Ct values up to 45 refer to the number of amplification cycles needed to detect the presence of the virus. These PCR results were compared with digital readings from the sensor to better evaluate the sensor technology. The results indicate that the samples with a range of Ct values from 17.8 to 35 can be differentiated, which highlights the increased sensitivity of this sensor technology. This research exhibits the potential of this biosensor technology to be further developed into a cost-effective, point-of-care, and portable rapid detection method for SARS-CoV-2.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）大流行在全球产生了重大影响。目前，该病毒最常见的检测方法是聚合酶链反应（PCR）和侧向流动检测。PCR需要一个多小时才能获得结果，而侧向流动检测在检测低浓度病毒时存在困难。在本研究中，使用一次性葡萄糖生物传感器条和可重复使用的印刷电路板，对60份临床人类唾液样本进行了新冠病毒（COVID-19）筛查，其中包括30份经逆转录聚合酶链反应（RT-PCR）确认的阳性样本和30份阴性样本。一次性试纸条进行了镀金和功能化处理，以便在金膜上固定抗体。功能化后，试纸条连接到印刷电路板上金属氧化物半导体场效应晶体管的栅极电极，以放大测试信号。向晶体管和试纸条的漏极施加同步双脉冲偏置电压。漏极波形的变化结果被转换为数字读数。使用定量PCR（RT-qPCR）再次检测经RT-PCR确认的唾液样本，以确定循环阈值（Ct）值。Ct值高达45表示检测病毒存在所需的扩增循环数。将这些PCR结果与传感器的数字读数进行比较，以更好地评估传感器技术。结果表明，Ct值在17.8至35范围内的样本可以区分，这突出了这种传感器技术灵敏度的提高。本研究展示了这种生物传感器技术进一步发展成为一种经济高效、即时检测和便携式的SARS-CoV-2快速检测方法的潜力。
