A novel G-quadruplex aptamer-based spike trimeric antigen test for the detection of SARS-CoV-2

Molecular Therapy: Nucleic Acids

Anbalagan Anantharaj¹  Guruprasad R. Medigeshi¹  Ankit Gupta²  Anjali Anand²  Tripti Shrivastava³  Neha Jain³  Sandeep Goswami⁴  Shinjini Bhatnagar⁴  Amit Kumar⁵  Rahul Singh⁶  Tarun Kumar Sharma⁶  Sharanabasava Patil⁶ 

[1] Technology Institute, NCR Biotech Science Cluster, Faridabad, Haryana 121001, India;Aptamer Technology and Diagnostics Laboratory, Multidisciplinary Clinical and Translational Research, Translational Health Science and Technology Institute, Faridabad, Haryana 121001, India;Discipline of Biosciences and Biomedical Engineering, Indian Institute of Technology Indore, Simrol, Madhya Pradesh 453552, India;;Infection and Immunology, Translational Health Science &Raja Ramanna Centre for Advanced Technology, Indore, Madhya Pradesh 452013, India;Translational Health Science and Technology Institute, Faridabad, Haryana 121001, India;

关键词: aptamer;    SARS-CoV-2;    diagnostics;    ALISA;    spike trimer;    batch-to-batch variation;   

DOI  :  

来源: DOAJ

【 摘 要 】

The recent SARS-CoV-2 outbreak has been declared a global health emergency. It will take years to vaccinate the whole population to protect them from this deadly virus, hence the management of SARS-CoV-2 largely depends on the widespread availability of an accurate diagnostic test. Toward addressing the unmet need of a reliable diagnostic test in the current work by utilizing the power of Systematic Evolution of Ligands by EXponential enrichment, a 44-mer G-quadruplex-forming DNA aptamer against spike trimer antigen of SARS-CoV-2 was identified. The lead aptamer candidate (S14) was characterized thoroughly for its binding, selectivity, affinity, structure, and batch-to-batch variability by utilizing various biochemical, biophysical, and in silico techniques. S14 has demonstrated a low nanomolar KD, confirming its tight binding to a spike antigen of SARS-CoV-2. S14 can detect as low as 2 nM of antigen. The clinical evaluation of S14 aptamer on nasopharyngeal swab specimens (n = 232) has displayed a highly discriminatory response between SARS-CoV-2 infected individuals from the non-infected one with a sensitivity and specificity of ∼91% and 98%, respectively. Importantly, S14 aptamer-based test has evinced a comparable performance with that of RT-PCR-based assay. Altogether, this study established the utility of aptamer technology for the detection of SARS-CoV-2.

【 授权许可】

Unknown