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

Ji Yeong Noh, Sun Woo Yoon, Youngji Kim, Thi Van Lo, Min Ju Ahn, Min Chul Jung, Tran Bac Le, Woonsung Na, Daesub Song, Van Phan Le, Seungjoo Haam, Dae Gwin Jeong, Hye Kwon Kim

Research output: Contribution to journalArticle

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.

Original languageEnglish
Article number16661
JournalScientific reports
Volume9
Issue number1
DOIs
Publication statusPublished - 2019 Dec 1

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Influenza A virus
Immunoassay
Point-of-Care Systems
Antigens
Limit of Detection
Color
Monoclonal Antibodies
Equipment and Supplies
Point-of-Care Testing
Antibodies
Proteins

All Science Journal Classification (ASJC) codes

  • General

Cite this

Noh, J. Y., Yoon, S. W., Kim, Y., Lo, T. V., Ahn, M. J., Jung, M. C., ... Kim, H. K. (2019). Pipetting-based immunoassay for point-of-care testing: Application for detection of the influenza A virus. Scientific reports, 9(1), [16661]. https://doi.org/10.1038/s41598-019-53083-8
Noh, Ji Yeong ; Yoon, Sun Woo ; Kim, Youngji ; Lo, Thi Van ; Ahn, Min Ju ; Jung, Min Chul ; Le, Tran Bac ; Na, Woonsung ; Song, Daesub ; Le, Van Phan ; Haam, Seungjoo ; Jeong, Dae Gwin ; Kim, Hye Kwon. / Pipetting-based immunoassay for point-of-care testing : Application for detection of the influenza A virus. In: Scientific reports. 2019 ; Vol. 9, No. 1.
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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.",
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Noh, JY, Yoon, SW, Kim, Y, Lo, TV, Ahn, MJ, Jung, MC, Le, TB, Na, W, Song, D, Le, VP, Haam, S, Jeong, DG & Kim, HK 2019, 'Pipetting-based immunoassay for point-of-care testing: Application for detection of the influenza A virus', Scientific reports, vol. 9, no. 1, 16661. https://doi.org/10.1038/s41598-019-53083-8

Pipetting-based immunoassay for point-of-care testing : Application for detection of the influenza A virus. / Noh, Ji Yeong; Yoon, Sun Woo; Kim, Youngji; Lo, Thi Van; Ahn, Min Ju; Jung, Min Chul; Le, Tran Bac; Na, Woonsung; Song, Daesub; Le, Van Phan; Haam, Seungjoo; Jeong, Dae Gwin; Kim, Hye Kwon.

In: Scientific reports, Vol. 9, No. 1, 16661, 01.12.2019.

Research output: Contribution to journalArticle

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T2 - Application for detection of the influenza A virus

AU - Noh, Ji Yeong

AU - Yoon, Sun Woo

AU - Kim, Youngji

AU - Lo, Thi Van

AU - Ahn, Min Ju

AU - Jung, Min Chul

AU - Le, Tran Bac

AU - Na, Woonsung

AU - Song, Daesub

AU - Le, Van Phan

AU - Haam, Seungjoo

AU - Jeong, Dae Gwin

AU - Kim, Hye Kwon

PY - 2019/12/1

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N2 - 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.

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