期刊
PARASITES & VECTORS
卷 11, 期 -, 页码 -出版社
BMC
DOI: 10.1186/s13071-018-2836-2
关键词
Leishmaniasis; Visceral leishmaniasis; Diagnostics; Loop-mediated isothermal amplification; LAMP; Less-invasive diagnosis; Non-invasive diagnosis; Loopamp (TM) Leishmania detection kit; Real-time fluorimeters; Boil & Spin
资金
- Federal Ministry of Education and Research, Germany (KfW) [202060457]
- Ministry of Foreign Affairs, Government of the Netherlands [22211]
- Red de Investigacion Cooperativa de Enfermedades Tropicales, Subprograma RETICS del Plan Estatal de I+D+I [RD16CIII/0003/0002]
- FEDER Una manera de hacer Europa funds
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.
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