Development of loop-mediated isothermal amplification (LAMP) assay using SYBR safe and gold-nanoparticle probe for detection of Leishmania in HIV patients

Abstract Asymptomatic leishmaniasis cases have continuously increased, especially among patients with HIV who are at risk to develop further symptoms of cutaneous and visceral leishmaniasis. Thus, early diagnosis using a simple, sensitive and reliable diagnostic assay is important because population...

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Autores principales:	Toon Ruang-areerate, Charanyarut Sukphattanaudomchoke, Thanyapit Thita, Saovanee Leelayoova, Phunlerd Piyaraj, Mathirut Mungthin, Patcharapan Suwannin, Duangporn Polpanich, Tienrat Tangchaikeeree, Kulachart Jangpatarapongsa, Kiattawee Choowongkomon, Suradej Siripattanapipong
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/be02e32bde264c10ba56e25e47f8ce2a
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spelling	oai:doaj.org-article:be02e32bde264c10ba56e25e47f8ce2a2021-12-02T17:47:36ZDevelopment of loop-mediated isothermal amplification (LAMP) assay using SYBR safe and gold-nanoparticle probe for detection of Leishmania in HIV patients10.1038/s41598-021-91540-52045-2322https://doaj.org/article/be02e32bde264c10ba56e25e47f8ce2a2021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-91540-5https://doaj.org/toc/2045-2322Abstract Asymptomatic leishmaniasis cases have continuously increased, especially among patients with HIV who are at risk to develop further symptoms of cutaneous and visceral leishmaniasis. Thus, early diagnosis using a simple, sensitive and reliable diagnostic assay is important because populations at risk mostly reside in rural communities where laboratory equipment is limited. In this study, the highly sensitive and selective determination of Leishmania infection in asymptomatic HIV patients was achieved using dual indicators (SYBR safe and gold-nanoparticle probe; AuNP-probe) in one-step LAMP method based on basic instruments. The assay can be simply evaluated under the naked eye due to clear interpretation of fluorescent emission of LAMP-SYBR safe dye-complex and colorimetric precipitate of specific AuNP-probes. The sensitivities and specificities of fluorescent SYBR safe dye and AuNP-probe indicators were equal, which were as high as 94.1 and 97.1%, respectively. Additionally, detection limits were 102 parasites/mL (0.0147 ng/µL), ten times more sensitivity than other related studies. To empower leishmaniasis surveillance, this inexpensive one-step SYBR safe and AuNP-LAMP assay is reliably fast and simple for field diagnostics to point-of-care settings, which can be set up in all levels of health care facilities including resource limited areas, especially in low to middle income countries.Toon Ruang-areerateCharanyarut SukphattanaudomchokeThanyapit ThitaSaovanee LeelayoovaPhunlerd PiyarajMathirut MungthinPatcharapan SuwanninDuangporn PolpanichTienrat TangchaikeereeKulachart JangpatarapongsaKiattawee ChoowongkomonSuradej SiripattanapipongNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Toon Ruang-areerate Charanyarut Sukphattanaudomchoke Thanyapit Thita Saovanee Leelayoova Phunlerd Piyaraj Mathirut Mungthin Patcharapan Suwannin Duangporn Polpanich Tienrat Tangchaikeeree Kulachart Jangpatarapongsa Kiattawee Choowongkomon Suradej Siripattanapipong Development of loop-mediated isothermal amplification (LAMP) assay using SYBR safe and gold-nanoparticle probe for detection of Leishmania in HIV patients
description	Abstract Asymptomatic leishmaniasis cases have continuously increased, especially among patients with HIV who are at risk to develop further symptoms of cutaneous and visceral leishmaniasis. Thus, early diagnosis using a simple, sensitive and reliable diagnostic assay is important because populations at risk mostly reside in rural communities where laboratory equipment is limited. In this study, the highly sensitive and selective determination of Leishmania infection in asymptomatic HIV patients was achieved using dual indicators (SYBR safe and gold-nanoparticle probe; AuNP-probe) in one-step LAMP method based on basic instruments. The assay can be simply evaluated under the naked eye due to clear interpretation of fluorescent emission of LAMP-SYBR safe dye-complex and colorimetric precipitate of specific AuNP-probes. The sensitivities and specificities of fluorescent SYBR safe dye and AuNP-probe indicators were equal, which were as high as 94.1 and 97.1%, respectively. Additionally, detection limits were 102 parasites/mL (0.0147 ng/µL), ten times more sensitivity than other related studies. To empower leishmaniasis surveillance, this inexpensive one-step SYBR safe and AuNP-LAMP assay is reliably fast and simple for field diagnostics to point-of-care settings, which can be set up in all levels of health care facilities including resource limited areas, especially in low to middle income countries.
format	article
author	Toon Ruang-areerate Charanyarut Sukphattanaudomchoke Thanyapit Thita Saovanee Leelayoova Phunlerd Piyaraj Mathirut Mungthin Patcharapan Suwannin Duangporn Polpanich Tienrat Tangchaikeeree Kulachart Jangpatarapongsa Kiattawee Choowongkomon Suradej Siripattanapipong
author_facet	Toon Ruang-areerate Charanyarut Sukphattanaudomchoke Thanyapit Thita Saovanee Leelayoova Phunlerd Piyaraj Mathirut Mungthin Patcharapan Suwannin Duangporn Polpanich Tienrat Tangchaikeeree Kulachart Jangpatarapongsa Kiattawee Choowongkomon Suradej Siripattanapipong
author_sort	Toon Ruang-areerate
title	Development of loop-mediated isothermal amplification (LAMP) assay using SYBR safe and gold-nanoparticle probe for detection of Leishmania in HIV patients
title_short	Development of loop-mediated isothermal amplification (LAMP) assay using SYBR safe and gold-nanoparticle probe for detection of Leishmania in HIV patients
title_full	Development of loop-mediated isothermal amplification (LAMP) assay using SYBR safe and gold-nanoparticle probe for detection of Leishmania in HIV patients
title_fullStr	Development of loop-mediated isothermal amplification (LAMP) assay using SYBR safe and gold-nanoparticle probe for detection of Leishmania in HIV patients
title_full_unstemmed	Development of loop-mediated isothermal amplification (LAMP) assay using SYBR safe and gold-nanoparticle probe for detection of Leishmania in HIV patients
title_sort	development of loop-mediated isothermal amplification (lamp) assay using sybr safe and gold-nanoparticle probe for detection of leishmania in hiv patients
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/be02e32bde264c10ba56e25e47f8ce2a
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Development of loop-mediated isothermal amplification (LAMP) assay using SYBR safe and gold-nanoparticle probe for detection of Leishmania in HIV patients

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