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A novel loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) device for rapid detection of Toxoplasma gondii in the blood of stray cats and dogs

Toxoplasma gondii is an obligate intracellular protozoan parasite that causes toxoplasmosis and threatens warm-blooded animal and human health worldwide. Simple and applicable diagnostic methods are urgently needed to guide development of effective approaches for prevention of toxoplasmosis. Most mo...

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Autores principales: Xue, Yangji, Kong, Qingming, Ding, Haojie, Xie, Chengzuo, Zheng, Bin, Zhuo, Xunhui, Ding, Jianzu, Tong, Qunbo, Lou, Di, Lu, Shaohong, Lv, Hangjun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: EDP Sciences 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8095094/
https://www.ncbi.nlm.nih.gov/pubmed/33944774
http://dx.doi.org/10.1051/parasite/2021039
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author Xue, Yangji
Kong, Qingming
Ding, Haojie
Xie, Chengzuo
Zheng, Bin
Zhuo, Xunhui
Ding, Jianzu
Tong, Qunbo
Lou, Di
Lu, Shaohong
Lv, Hangjun
author_facet Xue, Yangji
Kong, Qingming
Ding, Haojie
Xie, Chengzuo
Zheng, Bin
Zhuo, Xunhui
Ding, Jianzu
Tong, Qunbo
Lou, Di
Lu, Shaohong
Lv, Hangjun
author_sort Xue, Yangji
collection PubMed
description Toxoplasma gondii is an obligate intracellular protozoan parasite that causes toxoplasmosis and threatens warm-blooded animal and human health worldwide. Simple and applicable diagnostic methods are urgently needed to guide development of effective approaches for prevention of toxoplasmosis. Most molecular diagnostic tools for T. gondii infection require high technical skills, sophisticated equipment, and a controlled lab environment. In this study, we developed a loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) assay that specifically targets the 529 bp for detecting T. gondii infection. This novel portable device is universal, fast, user-friendly, and guarantees experimental sensitivity as well as low risk of aerosol contamination. Our LAMP-LFD assay has a detection limit of 1 fg of T. gondii DNA, and shows no cross-reaction with other parasitic pathogens, including Cryptosporidium parvum, Leishmania donovani, and Plasmodium vivax. We validated the developed assay by detecting T. gondii in DNA extracted from blood samples collected from 318 stray cats and dogs sampled from Deqing, Wenzhou, Yiwu, Lishui and Zhoushan cities across Zhejiang province, Eastern China. The LAMP-LFD device detected T. gondii DNA in 4.76 and 4.69% of stray cats and dogs, respectively. In conclusion, the developed LAMP-LFD assay is efficient, minimizes aerosol contamination, and is therefore suitable for detecting T. gondii across basic medical institutions and field settings.
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spelling pubmed-80950942021-05-06 A novel loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) device for rapid detection of Toxoplasma gondii in the blood of stray cats and dogs Xue, Yangji Kong, Qingming Ding, Haojie Xie, Chengzuo Zheng, Bin Zhuo, Xunhui Ding, Jianzu Tong, Qunbo Lou, Di Lu, Shaohong Lv, Hangjun Parasite Research Article Toxoplasma gondii is an obligate intracellular protozoan parasite that causes toxoplasmosis and threatens warm-blooded animal and human health worldwide. Simple and applicable diagnostic methods are urgently needed to guide development of effective approaches for prevention of toxoplasmosis. Most molecular diagnostic tools for T. gondii infection require high technical skills, sophisticated equipment, and a controlled lab environment. In this study, we developed a loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) assay that specifically targets the 529 bp for detecting T. gondii infection. This novel portable device is universal, fast, user-friendly, and guarantees experimental sensitivity as well as low risk of aerosol contamination. Our LAMP-LFD assay has a detection limit of 1 fg of T. gondii DNA, and shows no cross-reaction with other parasitic pathogens, including Cryptosporidium parvum, Leishmania donovani, and Plasmodium vivax. We validated the developed assay by detecting T. gondii in DNA extracted from blood samples collected from 318 stray cats and dogs sampled from Deqing, Wenzhou, Yiwu, Lishui and Zhoushan cities across Zhejiang province, Eastern China. The LAMP-LFD device detected T. gondii DNA in 4.76 and 4.69% of stray cats and dogs, respectively. In conclusion, the developed LAMP-LFD assay is efficient, minimizes aerosol contamination, and is therefore suitable for detecting T. gondii across basic medical institutions and field settings. EDP Sciences 2021-05-03 /pmc/articles/PMC8095094/ /pubmed/33944774 http://dx.doi.org/10.1051/parasite/2021039 Text en © Y. Xue et al., published by EDP Sciences, 2021 https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0 (https://creativecommons.org/licenses/by/4.0/) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Xue, Yangji
Kong, Qingming
Ding, Haojie
Xie, Chengzuo
Zheng, Bin
Zhuo, Xunhui
Ding, Jianzu
Tong, Qunbo
Lou, Di
Lu, Shaohong
Lv, Hangjun
A novel loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) device for rapid detection of Toxoplasma gondii in the blood of stray cats and dogs
title A novel loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) device for rapid detection of Toxoplasma gondii in the blood of stray cats and dogs
title_full A novel loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) device for rapid detection of Toxoplasma gondii in the blood of stray cats and dogs
title_fullStr A novel loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) device for rapid detection of Toxoplasma gondii in the blood of stray cats and dogs
title_full_unstemmed A novel loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) device for rapid detection of Toxoplasma gondii in the blood of stray cats and dogs
title_short A novel loop-mediated isothermal amplification-lateral-flow-dipstick (LAMP-LFD) device for rapid detection of Toxoplasma gondii in the blood of stray cats and dogs
title_sort novel loop-mediated isothermal amplification-lateral-flow-dipstick (lamp-lfd) device for rapid detection of toxoplasma gondii in the blood of stray cats and dogs
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8095094/
https://www.ncbi.nlm.nih.gov/pubmed/33944774
http://dx.doi.org/10.1051/parasite/2021039
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