Implementation of microfluidic sandwich ELISA for superior detection of plant pathogens.

Rapid and economical screening of plant pathogens is a high-priority need in the seed industry. Crop quality control and disease surveillance demand early and accurate detection in addition to robustness, scalability, and cost efficiency typically required for selective breeding and certification pr...

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Autores principales:	Numrin Thaitrong, Ratthaphol Charlermroj, Orawan Himananto, Channarong Seepiban, Nitsara Karoonuthaisiri
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f7a407ab06c14384acbf72b19dd1f798
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spelling	oai:doaj.org-article:f7a407ab06c14384acbf72b19dd1f7982021-11-18T08:40:38ZImplementation of microfluidic sandwich ELISA for superior detection of plant pathogens.1932-620310.1371/journal.pone.0083231https://doaj.org/article/f7a407ab06c14384acbf72b19dd1f7982013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24376668/?tool=EBIhttps://doaj.org/toc/1932-6203Rapid and economical screening of plant pathogens is a high-priority need in the seed industry. Crop quality control and disease surveillance demand early and accurate detection in addition to robustness, scalability, and cost efficiency typically required for selective breeding and certification programs. Compared to conventional bench-top detection techniques routinely employed, a microfluidic-based approach offers unique benefits to address these needs simultaneously. To our knowledge, this work reports the first attempt to perform microfluidic sandwich ELISA for Acidovorax citrulli (Ac), watermelon silver mottle virus (WSMoV), and melon yellow spot virus (MYSV) screening. The immunoassay occurs on the surface of a reaction chamber represented by a microfluidic channel. The capillary force within the microchannel draws a reagent into the reaction chamber as well as facilitates assay incubation. Because the underlying pad automatically absorbs excess fluid, the only operation required is sequential loading of buffers/reagents. Buffer selection, antibody concentrations, and sample loading scheme were optimized for each pathogen. Assay optimization reveals that the 20-folds lower sample volume demanded by the microchannel structure outweighs the 2- to 4-folds higher antibody concentrations required, resulting in overall 5-10 folds of reagent savings. In addition to cutting the assay time by more than 50%, the new platform offers 65% cost savings from less reagent consumption and labor cost. Our study also shows 12.5-, 2-, and 4-fold improvement in assay sensitivity for Ac, WSMoV, and MYSV, respectively. Practical feasibility is demonstrated using 19 real plant samples. Given a standard 96-well plate format, the developed assay is compatible with commercial fluorescent plate readers and readily amendable to robotic liquid handling systems for completely hand-free assay automation.Numrin ThaitrongRatthaphol CharlermrojOrawan HimanantoChannarong SeepibanNitsara KaroonuthaisiriPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 12, p e83231 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Numrin Thaitrong Ratthaphol Charlermroj Orawan Himananto Channarong Seepiban Nitsara Karoonuthaisiri Implementation of microfluidic sandwich ELISA for superior detection of plant pathogens.
description	Rapid and economical screening of plant pathogens is a high-priority need in the seed industry. Crop quality control and disease surveillance demand early and accurate detection in addition to robustness, scalability, and cost efficiency typically required for selective breeding and certification programs. Compared to conventional bench-top detection techniques routinely employed, a microfluidic-based approach offers unique benefits to address these needs simultaneously. To our knowledge, this work reports the first attempt to perform microfluidic sandwich ELISA for Acidovorax citrulli (Ac), watermelon silver mottle virus (WSMoV), and melon yellow spot virus (MYSV) screening. The immunoassay occurs on the surface of a reaction chamber represented by a microfluidic channel. The capillary force within the microchannel draws a reagent into the reaction chamber as well as facilitates assay incubation. Because the underlying pad automatically absorbs excess fluid, the only operation required is sequential loading of buffers/reagents. Buffer selection, antibody concentrations, and sample loading scheme were optimized for each pathogen. Assay optimization reveals that the 20-folds lower sample volume demanded by the microchannel structure outweighs the 2- to 4-folds higher antibody concentrations required, resulting in overall 5-10 folds of reagent savings. In addition to cutting the assay time by more than 50%, the new platform offers 65% cost savings from less reagent consumption and labor cost. Our study also shows 12.5-, 2-, and 4-fold improvement in assay sensitivity for Ac, WSMoV, and MYSV, respectively. Practical feasibility is demonstrated using 19 real plant samples. Given a standard 96-well plate format, the developed assay is compatible with commercial fluorescent plate readers and readily amendable to robotic liquid handling systems for completely hand-free assay automation.
format	article
author	Numrin Thaitrong Ratthaphol Charlermroj Orawan Himananto Channarong Seepiban Nitsara Karoonuthaisiri
author_facet	Numrin Thaitrong Ratthaphol Charlermroj Orawan Himananto Channarong Seepiban Nitsara Karoonuthaisiri
author_sort	Numrin Thaitrong
title	Implementation of microfluidic sandwich ELISA for superior detection of plant pathogens.
title_short	Implementation of microfluidic sandwich ELISA for superior detection of plant pathogens.
title_full	Implementation of microfluidic sandwich ELISA for superior detection of plant pathogens.
title_fullStr	Implementation of microfluidic sandwich ELISA for superior detection of plant pathogens.
title_full_unstemmed	Implementation of microfluidic sandwich ELISA for superior detection of plant pathogens.
title_sort	implementation of microfluidic sandwich elisa for superior detection of plant pathogens.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/f7a407ab06c14384acbf72b19dd1f798
work_keys_str_mv	AT numrinthaitrong implementationofmicrofluidicsandwichelisaforsuperiordetectionofplantpathogens AT ratthapholcharlermroj implementationofmicrofluidicsandwichelisaforsuperiordetectionofplantpathogens AT orawanhimananto implementationofmicrofluidicsandwichelisaforsuperiordetectionofplantpathogens AT channarongseepiban implementationofmicrofluidicsandwichelisaforsuperiordetectionofplantpathogens AT nitsarakaroonuthaisiri implementationofmicrofluidicsandwichelisaforsuperiordetectionofplantpathogens
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Implementation of microfluidic sandwich ELISA for superior detection of plant pathogens.

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