Detection of Streptococcus equi subsp. equi in guttural pouch lavage samples using a loop‐mediated isothermal nucleic acid amplification microfluidic device

Abstract Background Rapid point‐of‐care (POC) detection of Streptococcus equi subsp. equi (S. equi) would theoretically reduce the spread of strangles by identifying index and carrier horses. Hypothesis That the eqbE isothermal amplification (LAMP) assay, and the same eqbE LAMP assay tested in a mic...

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Autores principales:	Ashley G. Boyle, Shelley C. Rankin, Kathleen O'Shea, Darko Stefanovski, Jing Peng, Jinzhao Song, Haim H. Bau
Formato:	article
Lenguaje:	EN
Publicado:	Wiley 2021
Materias:	diagnostics DNA amplification equine point‐of‐care strangles Streptococcus equi Veterinary medicine SF600-1100
Acceso en línea:	https://doaj.org/article/72f9f71e37bd4ec7b43fcd27b2f3feba
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spelling	oai:doaj.org-article:72f9f71e37bd4ec7b43fcd27b2f3feba2021-11-30T17:01:05ZDetection of Streptococcus equi subsp. equi in guttural pouch lavage samples using a loop‐mediated isothermal nucleic acid amplification microfluidic device1939-16760891-664010.1111/jvim.16105https://doaj.org/article/72f9f71e37bd4ec7b43fcd27b2f3feba2021-05-01T00:00:00Zhttps://doi.org/10.1111/jvim.16105https://doaj.org/toc/0891-6640https://doaj.org/toc/1939-1676Abstract Background Rapid point‐of‐care (POC) detection of Streptococcus equi subsp. equi (S. equi) would theoretically reduce the spread of strangles by identifying index and carrier horses. Hypothesis That the eqbE isothermal amplification (LAMP) assay, and the same eqbE LAMP assay tested in a microfluidic device format, are comparable to a triplex real‐time quantitative polymerase chain reaction (qPCR) assay that is commonly used in diagnostic labs. Samples Sixty‐eight guttural pouch lavage (GPL) specimens from horses recovering from strangles. Methods Guttural pouch lavage specimens were tested for S. equi retrospectively using the benchtop eqbE LAMP, the eqbE LAMP microfluidic device, and compared to the triplex qPCR, that detects 2 S. equi‐specific genes, eqbE and SEQ2190, as the reference standard using the receiver operating characteristic area under the curve (ROC). Results The 27/68 specimens were positive by benchtop eqbE LAMP, 31/64 by eqbE LAMP microfluidic device, and 12/67 by triplex qPCR. Using the triplex PCR as the reference, the benchtop eqbE LAMP showed excellent discrimination (ROC Area = 0.813, 95% confidence interval [CI] = 0.711‐0.915) as did the LAMP microfluidic device (ROC Area = 0.811, 95% CI = 0.529‐0.782). There was no significant difference between the benchtop LAMP and LAMP microfluidic device (ROC Area 0.813 ± 0.055 vs 0.811 ± 0.034, P = .97). Conclusions The eqbE LAMP microfluidic device detected S. equi in GPL specimens from convalescent horses. This assay shows potential for development as a POC device for rapid, sensitive, accurate, and cost‐efficient detection of S. equi.Ashley G. BoyleShelley C. RankinKathleen O'SheaDarko StefanovskiJing PengJinzhao SongHaim H. BauWileyarticlediagnosticsDNA amplificationequinepoint‐of‐carestranglesStreptococcus equiVeterinary medicineSF600-1100ENJournal of Veterinary Internal Medicine, Vol 35, Iss 3, Pp 1597-1603 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	diagnostics DNA amplification equine point‐of‐care strangles Streptococcus equi Veterinary medicine SF600-1100
spellingShingle	diagnostics DNA amplification equine point‐of‐care strangles Streptococcus equi Veterinary medicine SF600-1100 Ashley G. Boyle Shelley C. Rankin Kathleen O'Shea Darko Stefanovski Jing Peng Jinzhao Song Haim H. Bau Detection of Streptococcus equi subsp. equi in guttural pouch lavage samples using a loop‐mediated isothermal nucleic acid amplification microfluidic device
description	Abstract Background Rapid point‐of‐care (POC) detection of Streptococcus equi subsp. equi (S. equi) would theoretically reduce the spread of strangles by identifying index and carrier horses. Hypothesis That the eqbE isothermal amplification (LAMP) assay, and the same eqbE LAMP assay tested in a microfluidic device format, are comparable to a triplex real‐time quantitative polymerase chain reaction (qPCR) assay that is commonly used in diagnostic labs. Samples Sixty‐eight guttural pouch lavage (GPL) specimens from horses recovering from strangles. Methods Guttural pouch lavage specimens were tested for S. equi retrospectively using the benchtop eqbE LAMP, the eqbE LAMP microfluidic device, and compared to the triplex qPCR, that detects 2 S. equi‐specific genes, eqbE and SEQ2190, as the reference standard using the receiver operating characteristic area under the curve (ROC). Results The 27/68 specimens were positive by benchtop eqbE LAMP, 31/64 by eqbE LAMP microfluidic device, and 12/67 by triplex qPCR. Using the triplex PCR as the reference, the benchtop eqbE LAMP showed excellent discrimination (ROC Area = 0.813, 95% confidence interval [CI] = 0.711‐0.915) as did the LAMP microfluidic device (ROC Area = 0.811, 95% CI = 0.529‐0.782). There was no significant difference between the benchtop LAMP and LAMP microfluidic device (ROC Area 0.813 ± 0.055 vs 0.811 ± 0.034, P = .97). Conclusions The eqbE LAMP microfluidic device detected S. equi in GPL specimens from convalescent horses. This assay shows potential for development as a POC device for rapid, sensitive, accurate, and cost‐efficient detection of S. equi.
format	article
author	Ashley G. Boyle Shelley C. Rankin Kathleen O'Shea Darko Stefanovski Jing Peng Jinzhao Song Haim H. Bau
author_facet	Ashley G. Boyle Shelley C. Rankin Kathleen O'Shea Darko Stefanovski Jing Peng Jinzhao Song Haim H. Bau
author_sort	Ashley G. Boyle
title	Detection of Streptococcus equi subsp. equi in guttural pouch lavage samples using a loop‐mediated isothermal nucleic acid amplification microfluidic device
title_short	Detection of Streptococcus equi subsp. equi in guttural pouch lavage samples using a loop‐mediated isothermal nucleic acid amplification microfluidic device
title_full	Detection of Streptococcus equi subsp. equi in guttural pouch lavage samples using a loop‐mediated isothermal nucleic acid amplification microfluidic device
title_fullStr	Detection of Streptococcus equi subsp. equi in guttural pouch lavage samples using a loop‐mediated isothermal nucleic acid amplification microfluidic device
title_full_unstemmed	Detection of Streptococcus equi subsp. equi in guttural pouch lavage samples using a loop‐mediated isothermal nucleic acid amplification microfluidic device
title_sort	detection of streptococcus equi subsp. equi in guttural pouch lavage samples using a loop‐mediated isothermal nucleic acid amplification microfluidic device
publisher	Wiley
publishDate	2021
url	https://doaj.org/article/72f9f71e37bd4ec7b43fcd27b2f3feba
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Detection of Streptococcus equi subsp. equi in guttural pouch lavage samples using a loop‐mediated isothermal nucleic acid amplification microfluidic device

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