2026-06-15T14:36:17Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/66412025-05-23T09:48:29Zcom_20.500.12105_2052com_20.500.12105_2051col_20.500.12105_19609
Repisalud
author
Ibarra-Meneses, Ana Victoria
author
Cruz, Israel
author
Chicharro, Carmen
author
Sánchez, Carmen
author
Biéler, Sylvain
author
Broger, Tobias
author
Moreno, Javier
author
Carrillo, Eugenia
funder
Federal Ministry of Education & Research (Alemania)
funder
Ministry of Foreign Affairs (Holanda)
2018-11-20T12:34:27Z
2018-11-20T12:34:27Z
2018-04-17
Parasit Vectors. 2018 Apr 17;11(1):250.
10.1186/s13071-018-2836-2
1756-3305
1756-3305
Parasites & vectors
29665825
http://hdl.handle.net/20.500.12105/6641
BACKGROUND: Nucleic acid amplification tests (NAATs) have proven to be advantageous in the diagnosis of leishmaniases, allowing sensitive diagnosis of: (i) cutaneous leishmaniasis in long duration lesions and (ii) visceral leishmaniasis using a less-invasive sample like peripheral blood, in opposition to tissue aspiration required for parasite demonstration by microscopy. Despite their benefits, the implementation of NAATs for leishmaniasis diagnosis at the point-of-care has not been achieved yet, mostly due to the complexity and logistical issues associated with PCR-based methods. METHODS: In this work, we have evaluated the performance of a ready-to-use loop-mediated isothermal amplification (LAMP) kit using two real time fluorimeters to amplify leishmanial DNA obtained by silica column-based and Boil & Spin protocols. RESULTS: The different approaches used to run and interpret the LAMP reactions showed a performance equivalent to PCR and real-time PCR, using spiked and clinical samples. The time to positivity obtained with real-time fluorimetry showed an excellent correlation with both Ct values and parasite load from real-time quantitative PCR. CONCLUSIONS: The results obtained open the possibility of using a highly stable, ready-to-use LAMP kit for the accurate diagnosis of leishmaniasis at the point-of-care. Furthermore, the feasibility of relating time to positivity, determined with a portable real-time fluorimeter, with the parasite burden could have a wider application in the management of leishmaniasis, such as in treatment efficacy monitoring or as a pharmacodynamics tool in clinical trials.
eng
Boil & Spin
Diagnostics
LAMP
Leishmaniasis
Less-invasive diagnosis
Loop-mediated isothermal amplification
Loopamp™ Leishmania detection kit
Non-invasive diagnosis
Real-time fluorimeters
Visceral leishmaniasis
Evaluation of fluorimetry and direct visualization to interpret results of a loop-mediated isothermal amplification kit to detect Leishmania DNA
research article
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EvaluationOfFluorimetryAnd_2018.pdf
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text/plain
EvaluationOfFluorimetryAnd_2018.pdf.txt