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Development of Label-Free Colorimetric Assay for MERS-CoV Using Gold Nanoparticles

구분
방역연계범부처감염병R&D사업
저자명
Han-bi Kim, Min-seon Park, Joon-ki Hwang, Jin-Hwa Kim, Doo-Ryeon Chung, Kyu-sung Lee, Min-hee Kang
학술지명
ACS Sensors
발표년월
2019-05-07
작성일
2023-11-13
조회수
157

1. RFP : 4분류 질환의 항원/항체 반응에 대한 간이키트 및 기기 개발

2. 해당분과 : 4-1-1 과제

3. 과제명 : 4분류 감염성 질환 현장진단용 표면증강 라만 다중 면역진단키트 및 광학 리더 개발

4. 연구책임자 : 주재범(중앙대학교)


[Abstract]

Worldwide outbreaks of infectious diseases necessitate the development of rapid and accurate diagnostic methods. Colorimetric assays are a representative tool to simply identify the target molecules in specimens through color changes of an indicator (e.g., nanosized metallic particle, and dye molecules). The detection method is used to conrm the presence of biomarkers visually and measure absorbance of the colored compounds at a specic wavelength. In this study, we propose a colorimetric assay based on an extended form of double-stranded DNA (dsDNA) self-assembly shielded gold nanoparticles (AuNPs) under positive electrolyte (e.g., 0.1 M MgCl2) for detection of Middle East respiratory syndrome coronavirus (MERS-CoV). This platform is able to verify the existence of viral molecules through a localized surface plasmon resonance (LSPR) shift and color changes of AuNPs in the UVvis wavelength range. We designed a pair of thiol-modied probes at either the 5end or 3end to organize complementary base pairs with upstream of the E protein gene (upE) and open reading frames (ORF) 1a on MERS-CoV. The dsDNA of the target and probes forms a disulde-induced long self-assembled complex, which protects AuNPs from salt-induced aggregation and transition of optical properties. This colorimetric assay could discriminate down to 1 pmol/μL of 30 bp MERS-CoV and further be adapted for convenient on-site detection of other infectious diseases, especially in resource-limited settings.