Evaluation of fluorimetry and direct visualization to interpret results of a loop-mediated isothermal amplification kit to detect Leishmania DNA.
Parasit Vectors
; 11(1): 250, 2018 04 17.
Article
em En
| MEDLINE
| ID: mdl-29665825
ABSTRACT
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.Palavras-chave
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Leishmaniose
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Técnicas de Amplificação de Ácido Nucleico
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Técnicas de Diagnóstico Molecular
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Fluorometria
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Leishmania
Tipo de estudo:
Diagnostic_studies
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Evaluation_studies
/
Guideline
Limite:
Humans
Idioma:
En
Revista:
Parasit Vectors
Ano de publicação:
2018
Tipo de documento:
Article
País de afiliação:
Espanha