Тип публикации: статья из журнала
Год издания: 2023
Идентификатор DOI: 10.1016/j.omtn.2023.02.010
Ключевые слова: MT: Oligonucleotides: Diagnostics and Biosensors COVID-19 diagnosis SARS-CoV-2 nucleocapsid detection label-free optical aptasensor aptamer selection biolayer interferometry binding motif identification
Аннотация: The spread of COVID-19 has affected billions of people across the globe, and the diagnosis of viral infection still needs improvement. Because of high immunogenicity and abundant expression during viral infection, SARS-CoV-2 nucleocapsid (N) protein could be an important diagnostic marker. This study aimed to develop a label-free oПоказать полностьюptical aptasensor fabricated with a novel single-stranded DNA aptamer to detect the N protein. The N-binding aptamers selected using asymmetric-emulsion PCR-SELEX and their binding affinity and cross-reactivity were characterized by biolayer interferometry. The tNSP3 aptamer (44 nt) was identified to bind the N protein of wild type and Delta and Omicron variants with high affinity (KD in the range of 0.6–3.5 nM). Utilizing tNSP3 to detect the N protein spiked in human saliva evinced the potential of this aptamer with a limit of detection of 4.5 nM. Mass spectrometry analysis was performed along with molecular dynamics simulation to obtain an insight into how tNSP3 binds to the N protein. The identified epitope peptides are localized within the RNA-binding domain and C terminus of the N protein. Hence, we confirmed the performance of this aptamer as an analytical tool for COVID-19 diagnosis.
Журнал: Molecular Therapy - Nucleic Acids
Выпуск журнала: Т.31
Номера страниц: 731-743
ISSN журнала: 21622531