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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doaj-be02e32bde264c10ba56e25e47f8ce2a2021-06-13T11:37:35ZengNature Publishing GroupScientific Reports2045-23222021-06-0111111110.1038/s41598-021-91540-5Development of loop-mediated isothermal amplification (LAMP) assay using SYBR safe and gold-nanoparticle probe for detection of Leishmania in HIV patientsToon Ruang-areerate0Charanyarut Sukphattanaudomchoke1Thanyapit Thita2Saovanee Leelayoova3Phunlerd Piyaraj4Mathirut Mungthin5Patcharapan Suwannin6Duangporn Polpanich7Tienrat Tangchaikeeree8Kulachart Jangpatarapongsa9Kiattawee Choowongkomon10Suradej Siripattanapipong11Department of Parasitology, Phramongkutklao College of MedicineDepartment of Microbiology, Faculty of Science, Mahidol UniversityDepartment of Parasitology, Phramongkutklao College of MedicineDepartment of Parasitology, Phramongkutklao College of MedicineDepartment of Parasitology, Phramongkutklao College of MedicineDepartment of Parasitology, Phramongkutklao College of MedicineCenter for Research and Innovation, Faculty of Medical Technology, Mahidol UniversityNational Nanotechnology Center (NANOTEC), National Science and Technology Development Agency (NSTDA)Center for Research and Innovation, Faculty of Medical Technology, Mahidol UniversityCenter for Research and Innovation, Faculty of Medical Technology, Mahidol UniversityDepartment of Biochemistry, Faculty of Science, Kasetsart UniversityDepartment of Microbiology, Faculty of Science, Mahidol UniversityAbstract 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.https://doi.org/10.1038/s41598-021-91540-5 |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
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 |
spellingShingle |
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 Scientific Reports |
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 Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2021-06-01 |
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. |
url |
https://doi.org/10.1038/s41598-021-91540-5 |
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