Dataset Information

A HiPAD Integrated with rGO/MWCNTs Nano-Circuit Heater for Visual Point-of-Care Testing of SARS-CoV-2.

ABSTRACT: Nowadays, the main obstacle for further miniaturization and integration of nucleic acids point-of-care testing devices is the lack of low-cost and high-performance heating materials for supporting reliable nucleic acids amplification. Herein, reduced graphene oxide hybridized multi-walled carbon nanotubes nano-circuit integrated into an ingenious paper-based heater is developed, which is integrated into a paper-based analytical device (named HiPAD). The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is still raging across the world. As a proof of concept, the HiPAD is utilized to visually detect the SARS-CoV-2 N gene using colored loop-mediated isothermal amplification reaction. This HiPAD costing a few dollars has comparable detection performance to traditional nucleic acids amplifier costing thousands of dollars. The detection range is from 25 to 2.5 × 10¹⁰ copies mL^-1 in 45 min. The detection limit of 25 copies mL^-1 is 40 times more sensitive than 1000 copies mL^-1 in conventional real-time PCR instruments. The disposable paper-based chip could also avoid potential secondary transmission of COVID-19 by convenient incineration to guarantee biosafety. The HiPAD or easily expanded M-HiPAD (for multiplex detection) has great potential for pathogen diagnostics in resource-limited settings.

SUBMITTER: Li S

PROVIDER: S-EPMC8250055 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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A HiPAD Integrated with rGO/MWCNTs Nano-Circuit Heater for Visual Point-of-Care Testing of SARS-CoV-2.

Li Sijie S Huang Shiyi S Ke Yuqing Y Chen Hongjun H Dang Jingqi J Huang Chengjie C Liu Wenjia W Cui Daxiang D Wang Jinglin J Zhi Xiao X Ding Xianting X

Advanced functional materials 20210420 26

Nowadays, the main obstacle for further miniaturization and integration of nucleic acids point-of-care testing devices is the lack of low-cost and high-performance heating materials for supporting reliable nucleic acids amplification. Herein, reduced graphene oxide hybridized multi-walled carbon nanotubes nano-circuit integrated into an ingenious paper-based heater is developed, which is integrated into a paper-based analytical device (named HiPAD). The coronavirus disease 2019 (COVID-19) pandem ...[more]

PMID: 34230825

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Project description:BackgroundThe COVID-19 pandemic has led to a rise in point-of-care (POC) and home-based tests, but concerns over usability, accuracy, and effectiveness have arisen. The incorporation of internal amplification controls (IACs), essential control for translational POC diagnostics, could mitigate false-negative and false-positive results due to sample matrix interference or inhibition. Although emerging POC nucleic acid amplification tests (NAATs) for detecting SARS-CoV-2 show impressive analytical sensitivity in the lab, the assessment of clinical accuracy with IACs is often overlooked. In some cases, the IACs were run spatially, complicating assay workflow. Therefore, the multiplex assay for pathogen and IAC is needed.ResultsWe developed a one-pot duplex reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay for saliva samples, a non-invasive and simple collected specimen for POC NAATs. The ORF1ab gene of SARS-CoV-2 was used as a target and a human 18S ribosomal RNA in human saliva was employed as an IAC to ensure clinical reliability of the RT-LAMP assay. The optimized assay could detect SARS-CoV-2 viral particles down to 100 copies/μL of saliva within 30 minutes without RNA extraction. The duplex RT-LAMP for SARS-CoV-2 and IAC is successfully amplified in the same reaction without cross-reactivity. The valid results were easily visualized in triple-line lateral flow immunoassay, in which two lines (flow control and IAC lines) represent valid negative results and three lines (flow control, IAC, and test line) represent valid positive results. This duplex assay demonstrated a clinical sensitivity of 95%, specificity of 100%, and accuracy of 96% in 30 clinical saliva samples.SignificanceIACs play a crucial role in ensuring user confidence with respect to the accuracy and reliability of at-home and POC molecular diagnostics. We demonstrated the multiplex capability of SARS-COV-2 and human18S ribosomal RNA RT-LAMP without complicating assay design. This generic platform can be extended in a similar manner to include human18S ribosomal RNA IACs into different clinical sample matrices.

Project description:BackgroundThe COVID-19 pandemic has led to a rise in point-of-care (POC) and home-based tests, but concerns over usability, accuracy, and effectiveness have arisen. The incorporation of internal amplification controls (IACs), essential control for translational POC diagnostics, could mitigate false-negative and false-positive results due to sample matrix interference or inhibition. Although emerging POC nucleic acid amplification tests (NAATs) for detecting SARS-CoV-2 show impressive analytical sensitivity in the lab, the assessment of clinical accuracy with IACs is often overlooked. In some cases, the IACs were run spatially, complicating assay workflow. Therefore, the multiplex assay for pathogen and IAC is needed.ResultsWe developed a one-pot duplex reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay for saliva samples, a non-invasive and simple collected specimen for POC NAATs. The ORF1ab gene of SARS-CoV-2 was used as a target and a human 18S ribosomal RNA in human saliva was employed as an IAC to ensure clinical reliability of the RT-LAMP assay. The optimized assay could detect SARS-CoV-2 viral particles down to 100 copies/μL of saliva within 30 min without RNA extraction. The duplex RT-LAMP for SARS-CoV-2 and IAC is successfully amplified in the same reaction without cross-reactivity. The valid results were easily visualized in triple-line lateral flow immunoassay, in which two lines (flow control and IAC lines) represent valid negative results and three lines (flow control, IAC, and test line) represent valid positive results. This duplex assay demonstrated a clinical sensitivity of 95%, specificity of 100%, and accuracy of 96% in 30 clinical saliva samples.SignificanceIACs play a crucial role in ensuring user confidence with respect to the accuracy and reliability of at-home and POC molecular diagnostics. We demonstrated the multiplex capability of SARS-COV-2 and human18S ribosomal RNA RT-LAMP without complicating assay design. This generic platform can be extended in a similar manner to include human18S ribosomal RNA IACs into different clinical sample matrices.

Dataset Information

A HiPAD Integrated with rGO/MWCNTs Nano-Circuit Heater for Visual Point-of-Care Testing of SARS-CoV-2.

Publications

A HiPAD Integrated with rGO/MWCNTs Nano-Circuit Heater for Visual Point-of-Care Testing of SARS-CoV-2.

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