Highly Sensitive and Specific Detection of Influenza A Viruses Using Bimolecular Fluorescence Complementation (BiFC) Reporter System

Author:

Lee Ui Jin1,Oh Yunkwang1,Kwon Oh Seok234,Shin Yong-Beom567ORCID,Kim Moonil157

Affiliation:

1. Critical Diseases Diagnostics Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 125 Gwahang-ro, Yuseong-gu, Daejeon 34141, Republic of Korea

2. SKKU Advanced Institute of Nanotechnology (SAINT), Sungkyunkwan University, Suwon 16419, Republic of Korea

3. Department of Nano Science and Technology, Sungkyunkwan University, Suwon 16419, Republic of Korea

4. Department of Nano Engineering, Sungkyunkwan University, Suwon 16419, Republic of Korea

5. Bionanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 34141, Republic of Korea

6. BioNano Health Guard Research Center (H-GUARD), Daejeon 34141, Republic of Korea

7. KRIBB School, Korea University of Science and Technology (UST), 217 Gajeong-ro, Yuseong-gu, Daejeon 34113, Republic of Korea

Abstract

In this study, we developed a highly sensitive and specific bimolecular fluorescence complementation (BiFC)-based influenza A virus (IAV)-sensing system by combining a galactose/glucose-binding protein (GGBP) with an N-terminal large domain (YN1-172) and a C-terminal small domain (YC173-239) made up of enhanced yellow fluorescence protein (eYFP). The GGBP-based BiFC reporter exhibits the fluorescence reconstitution as a result of conformational changes in GGBP when lactose, which was derived from 6′-silalyllactose and used as a substrate for neuraminidase (NA), binds to GGBP in the presence of IAV. The system showed a linear dynamic range extending from 1 × 100 to 1 × 107 TCID50/mL, and it had a detection limit of 1.1 × 100 TCID50/mL for IAV (H1N1), demonstrating ultra-high sensitivity. Our system exhibited fluorescence intensity enhancements in the presence of IAV, while it displayed weak fluorescence signals when exposed to NA-deficient viruses, such as RSV A, RSV B, adenovirus and rhinovirus, thereby indicating selective responses for IAV detection. Overall, our system provides a simple, highly sensitive and specific IAV detection platform based on BiFC that is capable of detecting ligand-induced protein conformational changes, obviating the need for virus culture or RNA extraction processes.

Funder

National Research Council of Science and Technology

Korea Research Institute of Bioscience and Biotechnology (KRIBB) Initiative Research Program

Bio and Medical Technology Development Program of the National Research Foundation

Publisher

MDPI AG

Subject

Clinical Biochemistry,General Medicine,Analytical Chemistry,Biotechnology,Instrumentation,Biomedical Engineering,Engineering (miscellaneous)

Reference46 articles.

1. A brief review of influenza virus infection;Javanian;J. Med. Virol.,2021

2. Influenza;Krammer;Nat. Rev. Dis. Primers,2018

3. Influenza vaccination and Guillain-Barré syndrome: Reality or fear;Babazadeh;J. Transl. Int. Med.,2019

4. The first influenza pandemic of the 21st century;Hajjar;Ann. Saudi. Med.,2010

5. Influenza virus: Dealing with a drifting and shifting pathogen;Kim;Viral Immunol.,2018

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3