Development of DNA Aptamers against the Nucleocapsid Protein of Severe Fever with Thrombocytopenia Syndrome Virus for Diagnostic Application: Catalytic Signal Amplification using Replication Protein A‑Conjugated Liposomes

Most prevalent infectious diseases worldwide are caused by mediators such as insects and characterized by high mortality and morbidity, thereby creating a global public health concern. Therefore, a sensitive, selective detection platform for diagnosing diseases in the early stages of infection is ne...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2019-11, Vol.91 (21), p.13772-13779
Main Authors: Yeom, Gyuho, Kang, Juyoung, Jang, Hyungjun, Nam, Ho Yeon, Kim, Min-Gon, Park, Chin-Ju
Format: Article
Language:English
Subjects:
Aptamers
Chemistry
Colorimetry
Cross-reactivity
Deoxyribonucleic acid
Diagnostic software
Diagnostic systems
Disease spread
DNA
Encapsulation
Fever
Horseradish peroxidase
Hydrogen peroxide
Infectious diseases
Insects
Liposomes
Lysis
Morbidity
Nucleocapsids
Peroxidase
Protein A
Proteins
Public health
Replication protein A
Thrombocytopenia
Viruses
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Summary:Most prevalent infectious diseases worldwide are caused by mediators such as insects and characterized by high mortality and morbidity, thereby creating a global public health concern. Therefore, a sensitive, selective detection platform for diagnosing diseases in the early stages of infection is needed to prevent disease spread and to protect public health. Here, we developed novel DNA aptamers specific to the nucleocapsid protein (NP) of the severe fever with thrombocytopenia syndrome (SFTS) virus and synthesized ssDNA-binding protein-conjugated liposomes encapsulated with horseradish peroxidase (HRP) for application in a simple and universal platform. This platform achieved highly sensitive detection of the NP by measuring the colorimetric signal following lysis of the HRP encapsulated liposomes, mediated by a mixture of 3,3′,5,5′-tetramethylbenzidine and H2O2 solution. The limit of detection was 0.009 ng·mL–1, and NP was successfully detected in diluted human serum with a high recovery rate. Moreover, this method was specific and did not exhibit cross-reactivity among NPs of other virus types. These results demonstrated the efficacy of the proposed method as a highly sensitive, specific, and universal diagnostic tool for potential application in monitoring of the early stages of infectious diseases.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.9b03210