Hijacking the self-replicating machine of bacteriophage for PCR-based cascade signal amplification in detecting SARS-CoV-2 viral marker protein in serum

Jialei Du; Daili Xiang; Fushan Liu; Leichen Wang; Hao Li; Liu Gong; Xiangyu Fan

doi:10.1016/j.snb.2022.132780

Hijacking the self-replicating machine of bacteriophage for PCR-based cascade signal amplification in detecting SARS-CoV-2 viral marker protein in serum

Sens Actuators B Chem. 2023 Jan 1:374:132780. doi: 10.1016/j.snb.2022.132780. Epub 2022 Oct 14.

Authors

Jialei Du¹, Daili Xiang¹, Fushan Liu¹, Leichen Wang¹, Hao Li^{1

2}, Liu Gong¹, Xiangyu Fan²

Affiliations

¹ Institute for Advanced Interdisciplinary Research (iAIR), Collaborative Innovation Center of Technology and Equipment for Biological Diagnosis and Therapy in Universities of Shandong, University of Jinan, Jinan 250022, China.
² School of Biological Science and Technology, University of Jinan, Jinan 250024, China.

Abstract

In this work, the nucleic acid detection of SARS-Cov-2 is extended to protein markers of the virus, utilizing bacteriophage. Specifically, the phage display technique enables the main protease of SARS-Cov-2 to control the self-replication of m13 phage, so that the presence of the viral protease can be amplified by phage replication as the first round of signal amplification. Then, the genome of replicated phage can be detected using polymer chain reaction (PCR), as the second round of signal amplification. Based on these two types of well-established biotechnology, the proposed method shows satisfactory sensitivity and robustness in the direct serum detection of the viral protease. These results may point to clinical application in the near future.

Keywords: Bacteriophage; COVID-19; Main protease; PCR; Phage display; SARS-Cov-2.