An alternative ready-to-use electrochemical immunosensor for point-of-care COVID-19 diagnosis using graphene screen-printed electrodes coupled with a 3D-printed portable potentiostat

•The modified graphene screen-printed electrode with poly(pyrrole propionic acid) and SARS-CoV-2 receptor binding domain (RBD) antibody was fabricated for COVID-19 detection by using in-house portable potentiostat.•Immunosensor based on enzymatic immunoassay was applied to determine quantitative of...

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Veröffentlicht in:Talanta open 2022-12, Vol.6, p.100155-100155, Article 100155
Hauptverfasser: Primpray, Vitsarut, Kamsong, Wichayaporn, Pakapongpan, Saithip, Phochakum, Kanchanok, Kaewchaem, Arissanan, Sappat, Assawapong, Wisitsoraat, Anurat, Lomas, Tanom, Tuantranont, Adisorn, Karuwan, Chanpen
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Sprache:eng
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Zusammenfassung:•The modified graphene screen-printed electrode with poly(pyrrole propionic acid) and SARS-CoV-2 receptor binding domain (RBD) antibody was fabricated for COVID-19 detection by using in-house portable potentiostat.•Immunosensor based on enzymatic immunoassay was applied to determine quantitative of SARS-CoV-2 receptor binding domain in nasopharyngeal samples.•The in-housed portable potentiostat was fabricated as a point of care testing device for the screening of COVID-19 infection. A severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a cause of worldwide Coronavirus 2019 (COVID-19) disease pandemic. It is thus important to develop ultra-sensitive, rapid and easy-to-use methods for the identification of COVID-19 infected patients. Herein, an alternative electrochemical immunosensor based on poly(pyrrolepropionic acid) (pPPA) modified graphene screen-printed electrode (GSPE) was proposed for rapid COVID-19 detection. The method was based on a competitive enzyme immunoassay process utilizing horseradish peroxidase (HRP)-conjugated SARS-CoV-2 as a reporter binding molecule to compete binding with antibody against the SARS-CoV-2 receptor binding domain (SARS-CoV-2 RBD) protein. This strategy enhanced the current signal via the enzymatic reaction of HRP-conjugated SARS-CoV-2 RBD antibody on the electrode surface. The modification, immobilization, blocking, and detection processes were optimized and evaluated by amperometry. The quantitative analysis of SARS-CoV-2 was conducted based on competitive enzyme immunoassay with amperometric detection using a 3D-printed portable potentiostat for point-of-care COVID-19 diagnosis. The current measurements at -0.2 V yielded a calibration curve with a linear range of 0.01–1500 ng mL−1 (r2 = 0.983), a low detection limit of 2 pg mL−1 and a low quantification limit of 10 pg mL−1. In addition, the analyzed results of practical samples using the developed method were successfully verified with ELISA and RT-PCR. Therefore, the proposed portable electrochemical immunosensor is highly sensitive, rapid, and reliable. Thus, it is an alternative ready-to-use sensor for COVID-19 point-of-care diagnosis. [Display omitted]
ISSN:2666-8319
2666-8319
DOI:10.1016/j.talo.2022.100155