Pipetting-based immunoassay for point-of-care testing: Application for detection of the influenza A virus

[tetano]

Sci Rep. 2019 Nov 13;9(1):16661. doi: 10.1038/s41598-019-53083-8. Pipetting-based immunoassay for point-of-care testing: Application for detection of the influenza A virus.

Noh JY^1,2, Yoon SW^2,3, Kim Y^2,4, Lo TV^2,3, Ahn MJ^2,3, Jung MC^2,3, Le TB^2,3, Na W^5,6, Song D⁵, Le VP⁷, Haam S⁸, Jeong DG^9,10, Kim HK^11,12.
Author information

1 Department of Microbiology, College of Natural Sciences, Chungbuk National University, Cheongju, Republic of Korea. 2 Infectious Diseases Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea. 3 Bio-Analytical Science Division, University of Science and Technology (UST), Daejeon, Republic of Korea. 4 College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea. 5 Department of Pharmacy, College of Pharmacy, Korea University, Sejong, Republic of Korea. 6 College of Veterinary Medicine, Chonnam National University, Gwangju, Korea. 7 Department of Microbiology and Infectious Diseases, College of Veterinary Medicine, Vietnam National University of Agriculture, Hanoi, Vietnam. 8 Department of Chemical and Biomolecular Engineering, Yonsei University, Yonsei-ro 50, Seoul, 03722, Republic of Korea. 9 Infectious Diseases Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea. dgjeong@kribb.re.kr. 10 Bio-Analytical Science Division, University of Science and Technology (UST), Daejeon, Republic of Korea. dgjeong@kribb.re.kr. 11 Department of Microbiology, College of Natural Sciences, Chungbuk National University, Cheongju, Republic of Korea. khk1329@chungbuk.ac.kr. 12 Infectious Diseases Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Republic of Korea. khk1329@chungbuk.ac.kr.

Abstract

Point-of-care tests (POCT) for pathogens are considered important for low-resource countries and facilities. Although lateral flow immunoassays (LFIA) have many advantages including speed and ease of use, their sensitivity is limited without specific equipment. Furthermore, their response cannot be enhanced through enzymatic reactions. Owing to these limitations, LFIAs have not yet been generally adopted as the standard protocol for in vitro analysis of infectious pathogens. We aimed to develop a novel pipetting-based immunoassay using a removable magnetic ring-coupled pipette tip. The "magnetic bead-capture antibody-targeted protein complex" was simply purified by pipetting and quantified by enzymatic colour development or using a lateral flow system. This pipetting-based immunoassay was applied to detect the nucleoprotein (NP) of the influenza A virus. Using an HRP-conjugated monoclonal antibody as a probe, the assay allowed for specific and sensitive detection. Furthermore, when this assay was applied exclusively for antigen capture in the lateral flow system, the limit of detection improved 100-fold and displayed greater sensitivity than the lateral flow system alone. Therefore, the pipetting-based immunoassay may be potentially used as a sensitive POCT to clinically detect a target antigen.


PMID: 31723156 DOI: 10.1038/s41598-019-53083-8