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Detection of SARS-CoV-2 N protein using AgNPs-modified aligned silicon nanowires BioSERS chip.

ABSTRACT: The SARS-CoV-2 (COVID-19) pandemic had a strong impact on societies and economies worldwide and tests for high-performance detection of SARS-CoV-2 biomarkers are still needed for potential future outbreaks of the disease. In this paper, we present the different steps for the design of an aptamer-based surface-enhanced Raman scattering (BioSERS) sensing chip capable of detecting the coronavirus nucleocapsid protein (N protein) in spiked phosphate-buffered solutions and real samples of human blood serum. Optimization of the preparation steps in terms of the aptamer concentration used for the functionalization of the silver nanoparticles, time for affixing the aptamer, incubation time with target protein, and insulation of the silver active surface with cysteamine, led to a sensitive BioSERS chip, which was able to detect the N protein in the range from 1 to 75 ng mL^-1 in spiked phosphate-buffered solutions with a detection limit of 1 ng mL^-1 within 30 min. Furthermore, the BioSERS chip was used to detect the target protein in scarcely spiked human serum. This study demonstrates the possibility of a clinical application that can improve the detection limit and accuracy of the currently commercialized SARS-CoV-2 immunodiagnostic kit. Additionally, the system is modular and can be applied to detect other proteins by only changing the aptamer.

SUBMITTER: Kouz S

PROVIDER: S-EPMC11019291 | biostudies-literature | 2024 Apr

REPOSITORIES: biostudies-literature

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Detection of SARS-CoV-2 N protein using AgNPs-modified aligned silicon nanowires BioSERS chip.

Kouz Sadok S Raouafi Amal A Ouhibi Awatef A Lorrain Nathalie N Essafi Makram M Mejri Manel M Raouafi Noureddine N Moadhen Adel A Guendouz Mohammed M

RSC advances 20240416 17

The SARS-CoV-2 (COVID-19) pandemic had a strong impact on societies and economies worldwide and tests for high-performance detection of SARS-CoV-2 biomarkers are still needed for potential future outbreaks of the disease. In this paper, we present the different steps for the design of an aptamer-based surface-enhanced Raman scattering (BioSERS) sensing chip capable of detecting the coronavirus nucleocapsid protein (N protein) in spiked phosphate-buffered solutions and real samples of human blood ...[more]

PMID: 38628480

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Project description:In 2019 the COVID-19 pandemic, caused by SARS-CoV-2, demonstrated the urgent need for rapid, reliable, and portable diagnostics. The COVID-19 pandemic was declared in January 2020 and surges of the outbreak continue to reoccur. It is clear that early identification of infected individuals, especially asymptomatic carriers, plays a huge role in preventing the spread of the disease. The current gold standard diagnostic for SARS-CoV-2 is quantitative reverse transcription polymerase chain reaction (qRT-PCR) test based on the detection of the viral RNA. While RT-PCR is reliable and sensitive, it requires expensive centralized equipment and is time consuming (∼2 h or more); limiting its applicability in low resource areas. The FDA issued Emergency Use Authorizations (EUAs) for several COVID-19 diagnostics with an emphasis on point-of care (PoC) testing. Numerous RT-PCR and serological tests were approved for use at the point of care. Abbott's ID NOW, and Cue Health's COVID-19 test are of particular interest, which use isothermal amplification methods for rapid detection in under 20 min. We look to expand on the range of current PoC testing platforms with a new rapid and portable isothermal nucleic acid detection device. We pair reverse transcription loop mediated isothermal amplification (RT-LAMP) with a particle imaging technique, particle diffusometry (PD), to successfully detect SARS-CoV-2 in only 35 min on a portable chip with integrated heating. A smartphone device is used to image the samples containing fluorescent beads post-RT-LAMP and correlates decreased diffusivity to positive samples. We detect as little as 30 virus particles per μL from a RT-LAMP reaction in a microfluidic chip using a portable heating unit. Further, we can perform RT-LAMP from a diluted unprocessed saliva sample without RNA extraction. Additionally, we lyophilize SARS-CoV-2-specific RT-LAMP reactions that target both the N gene and the ORF1ab gene in the microfluidic chip, eliminating the need for cold storage. Our assay meets specific target product profiles outlined by the World Health Organization: it is specific to SARS-CoV-2, does not require cold storage, is compatible with digital connectivity, and has a detection limit of less than 35 × 104 viral particles per mL in saliva. PD-LAMP is rapid, simple, and attractive for screening and use at the point of care.

Project description:Severe acute respiratory syndrome SARS-CoV-2 virus led to notable challenges amongst researchers in view of development of new and fast detecting techniques. In this regard, surface-enhanced Raman spectroscopy (SERS) technique, providing a fingerprint characteristic for each material, would be an interesting approach. The current study encompasses the fabrication of a SERS sensor to study the SARS-CoV-2 S1 (RBD) spike protein of the SARS-CoV-2 virus family. The SERS sensor consists of a silicon nanowires (SiNWs) substrate decorated with plasmonic silver nanoparticles (AgNPs). Both SiNWs fabrication and AgNPs decoration were achieved by a relatively simple wet chemical processing method. The study deliberately projects the factors that influence the growth of silicon nanowires, uniform decoration of AgNPs onto the SiNWs matrix along with detection of Rhodamine-6G (R6G) to optimize the best conditions for enhanced sensing of the spike protein. Increasing the time period of etching process resulted in enhanced SiNWs' length from 0.55 to 7.34 µm. Furthermore, the variation of the immersion time in the decoration process of AgNPs onto SiNWs ensued the optimum time period for the enhancement in the sensitivity of detection. Tremendous increase in sensitivity of R6G detection was perceived on SiNWs etched for 2 min (length=0.90 µm), followed by 30s of immersion time for their optimal decoration by AgNPs. These SiNWs/AgNPs SERS-based sensors were able to detect the spike protein at a concentration down to 9.3 × 10-12 M. Strong and dominant peaks at 1280, 1404, 1495, 1541 and 1609 cm-1 were spotted at a fraction of a minute. Moreover, direct, ultra-fast, facile, and affordable optoelectronic SiNWs/AgNPs sensors tuned to function as a biosensor for detecting the spike protein even at a trace level (pico molar concentration). The current findings hold great promise for the utilization of SERS as an innovative approach in the diagnosis domain of infections at very early stages.

Dataset Information

Detection of SARS-CoV-2 N protein using AgNPs-modified aligned silicon nanowires BioSERS chip.

Publications

Detection of SARS-CoV-2 N protein using AgNPs-modified aligned silicon nanowires BioSERS chip.

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