Validation and application of a visual LAMP assay for Mpox diagnosis — Insights from clinical samples in Thailand

Despite numerous recent advances in infectious disease diagnostic technologies including quantitative (q) polymerase chain reaction (PCR) and antigen test kit (ATK), there remains a need to develop more efficient point-of-care assays for large-scale operations. In an effort towards such a screening...

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Main Authors: Wansadaj Jaroenram, Yada Ongchanchai, Ravee Nitiyanontakij, Pawita Suwanwatthana, Ratchana Ponang, Wansika Kiatpathomchai
Format: Article
Language:English
Published: Elsevier 2025-12-01
Series:MethodsX
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Online Access:http://www.sciencedirect.com/science/article/pii/S2215016125004121
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author Wansadaj Jaroenram
Yada Ongchanchai
Ravee Nitiyanontakij
Pawita Suwanwatthana
Ratchana Ponang
Wansika Kiatpathomchai
author_facet Wansadaj Jaroenram
Yada Ongchanchai
Ravee Nitiyanontakij
Pawita Suwanwatthana
Ratchana Ponang
Wansika Kiatpathomchai
author_sort Wansadaj Jaroenram
collection DOAJ
description Despite numerous recent advances in infectious disease diagnostic technologies including quantitative (q) polymerase chain reaction (PCR) and antigen test kit (ATK), there remains a need to develop more efficient point-of-care assays for large-scale operations. In an effort towards such a screening test for low-resource settings, we have established a colorimetric loop-mediated isothermal amplification assay and detected the amplified DNA via naked-eye observation to maximize assay simplicity using monkeypox (Mpox) as a model. The assay is cheap, highly sensitive and specific to Mpox with the potential for translation into broader applications. • Our protocol offers a simpler, faster, and cheaper test for Mpox than the current gold-standard qPCR, with 100% accuracy. • assay enables decentralized screening and provides visually interpretable results immediately after incubation • The versatility of this protocol allows for easy adaptation to detecting new or re-emerging diseases, utilizing a visual readout for maximum simplicity.
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institution Kabale University
issn 2215-0161
language English
publishDate 2025-12-01
publisher Elsevier
record_format Article
series MethodsX
spelling doaj-art-b7173f519c6e433c90e736b9a00fe1e12025-08-24T05:12:48ZengElsevierMethodsX2215-01612025-12-011510356810.1016/j.mex.2025.103568Validation and application of a visual LAMP assay for Mpox diagnosis — Insights from clinical samples in ThailandWansadaj Jaroenram0Yada Ongchanchai1Ravee Nitiyanontakij2Pawita Suwanwatthana3Ratchana Ponang4Wansika Kiatpathomchai5National Center for Genetic Engineering and Biotechnology (BIOTEC), 113 Thailand Science Park, Phahonyothin Rd., Klong Neung, Klong Luang, Pathum Thani 1212, Thailand; Corresponding author.National Center for Genetic Engineering and Biotechnology (BIOTEC), 113 Thailand Science Park, Phahonyothin Rd., Klong Neung, Klong Luang, Pathum Thani 1212, Thailand; Faculty of Veterinary Technology, Kasetsart University, Bangkok 10900, ThailandBamrasnaradura Infectious Diseases Institute, 126 Tivanon Rd., Talat Khwan, Mueang, Nonthaburi 11000, ThailandBamrasnaradura Infectious Diseases Institute, 126 Tivanon Rd., Talat Khwan, Mueang, Nonthaburi 11000, ThailandBamrasnaradura Infectious Diseases Institute, 126 Tivanon Rd., Talat Khwan, Mueang, Nonthaburi 11000, ThailandNational Center for Genetic Engineering and Biotechnology (BIOTEC), 113 Thailand Science Park, Phahonyothin Rd., Klong Neung, Klong Luang, Pathum Thani 1212, ThailandDespite numerous recent advances in infectious disease diagnostic technologies including quantitative (q) polymerase chain reaction (PCR) and antigen test kit (ATK), there remains a need to develop more efficient point-of-care assays for large-scale operations. In an effort towards such a screening test for low-resource settings, we have established a colorimetric loop-mediated isothermal amplification assay and detected the amplified DNA via naked-eye observation to maximize assay simplicity using monkeypox (Mpox) as a model. The assay is cheap, highly sensitive and specific to Mpox with the potential for translation into broader applications. • Our protocol offers a simpler, faster, and cheaper test for Mpox than the current gold-standard qPCR, with 100% accuracy. • assay enables decentralized screening and provides visually interpretable results immediately after incubation • The versatility of this protocol allows for easy adaptation to detecting new or re-emerging diseases, utilizing a visual readout for maximum simplicity.http://www.sciencedirect.com/science/article/pii/S2215016125004121Colorimetric LAMPMonkeypox virusMpoxXylenol orangeXO
spellingShingle Wansadaj Jaroenram
Yada Ongchanchai
Ravee Nitiyanontakij
Pawita Suwanwatthana
Ratchana Ponang
Wansika Kiatpathomchai
Validation and application of a visual LAMP assay for Mpox diagnosis — Insights from clinical samples in Thailand
MethodsX
Colorimetric LAMP
Monkeypox virus
Mpox
Xylenol orange
XO
title Validation and application of a visual LAMP assay for Mpox diagnosis — Insights from clinical samples in Thailand
title_full Validation and application of a visual LAMP assay for Mpox diagnosis — Insights from clinical samples in Thailand
title_fullStr Validation and application of a visual LAMP assay for Mpox diagnosis — Insights from clinical samples in Thailand
title_full_unstemmed Validation and application of a visual LAMP assay for Mpox diagnosis — Insights from clinical samples in Thailand
title_short Validation and application of a visual LAMP assay for Mpox diagnosis — Insights from clinical samples in Thailand
title_sort validation and application of a visual lamp assay for mpox diagnosis insights from clinical samples in thailand
topic Colorimetric LAMP
Monkeypox virus
Mpox
Xylenol orange
XO
url http://www.sciencedirect.com/science/article/pii/S2215016125004121
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