Ultra-sensitive detection of SARS-CoV-2 S1 protein by coupling rolling circle amplification with poly(N-isopropylacrylamide)-based sandwich-type assay.

Talanta

College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, China; Department of Radiation and Medical Oncology, Hubei Key Laboratory of Tumor Biological Behaviors, and Hubei Cancer Clinical Study Center, Zhongnan Hospital of Wuhan University, Wuhan, 430072, China. Electronic addre

Published: November 2024


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Article Abstract

In the past few years, the COVID-19 pandemic, caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) seriously threatens global public health security due to its high contagiousness. It remains of vital importance to develop a rapid and sensitive assay for SARS-CoV-2. In this work, we proposed a sandwich-type assay based on poly(N-isopropylacrylamide) (PNIPAM), allowing efficient detection of the SARS-CoV-2 S1 protein in the homogeneous solution. Firstly, a direct sandwich-type assay was established with a linear range of 0.2-2 μg/mL and a limit of detection (LOD) of 0.11 μg/mL, which could realize rapid detection in about 1 h. Furthermore, the sandwich-type assay coupled with rolling circle amplification (RCA) obtained an increase in sensitivity of 5.9 × 10 folds with a wide linear range of 0.01 - 100 ng/mL and a LOD of 1.88 pg/mL. The average recoveries in unpretreated saliva were 90 %-113.0 %, indicating the potential of the developed method for application in practical samples. Given the high selectivity and sensitivity of the developed method, it has a significant potential for rapid and early detection of SARS-CoV-2.

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http://dx.doi.org/10.1016/j.talanta.2024.126572DOI Listing

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