Development of loop-mediated isothermal amplification (LAMP) for simple detection of Leishmania infection
Chaichontat Sriworarat,Atchara Phumee,Mathirut Mungthin,Saovanee Leelayoova,Padet Siriyasatien,Padet Siriyasatien +5 more
TLDR
A simple colorimetric loop-mediated isothermal amplification (LAMP) technique for the direct detection of Leishmania DNA could enable the rapid diagnosis of leishmaniasis, thereby facilitating the survey and control of leishingmaniasis in Thailand.Abstract:
Leishmaniasis is a neglected tropical disease that is caused by an obligate intracellular protozoan of the genus Leishmania. Recently, an increasing number of autochthonous leishmaniasis cases caused by L. martiniquensis and the novel species L. siamensis have been described in Thailand, rendering an accurate diagnosis of this disease critical. However, only a few laboratories are capable of diagnosing leishmaniasis in Thailand. To expand leishmaniasis diagnostic capabilities, we developed a simple colorimetric loop-mediated isothermal amplification (LAMP) technique for the direct detection of Leishmania DNA. LAMP was performed for 75 min using four primers targeting the conserved region of the18S ribosomal RNA gene, and the DNA indicator used was malachite green (MG). To simulate crude samples, cultured promastigotes of L. siamensis were mixed with blood or saliva. Also, clinical samples (blood, saliva, and tissue biopsies) were obtained from patients with cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL). All samples were boiled for 10 min and introduced directly into the LAMP reaction mixture without DNA purification. The use of MG resulted in an unambiguous differentiation of positive and negative controls. For L. siamensis, the detection limit was 103 parasites/mL or 2.5 parasites/tube. Saliva, tissue biopsies, and whole blood were indicative of active Leishmania infection, and their direct usages did not adversely affect the detection limit. In addition, this LAMP assay could detect DNA from multiple Leishmania species other than L. siamensis and L. martiniquensis, including L. aethiopica, L. braziliensis, L. donovani and L. tropica. The simplicity and sensitivity of LAMP in detecting active Leishmania infection could enable the rapid diagnosis of leishmaniasis, thereby facilitating the survey and control of leishmaniasis in Thailand. However, our limited number of samples warranted a further validation with a larger cohort of patients before this assay could be deployed.read more
Citations
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Journal ArticleDOI
Loop-mediated isothermal amplification (LAMP): a versatile technique for detection of micro-organisms.
TL;DR: Available types of LAMP techniques will be discussed together with their applications in detection of various micro‐organisms, and advantages and limitations of this molecular detection method are described together with its future potential as a diagnostic method for infectious disease.
Journal ArticleDOI
Leishmania infections: Molecular targets and diagnosis
Mohammad Akhoundi,Tim Downing,Jan Votýpka,Katrin Kuhls,Julius Lukeš,Arnaud Cannet,Christophe Ravel,Pierre Marty,Pascal Delaunay,Mohamed Kasbari,Bruno Granouillac,Luigi Gradoni,Denis Sereno +12 more
TL;DR: A classification method based on the suitability of well-studied molecular markers for typing the 21 known Leishmania species pathogenic to humans is proposed, which can be applied to newly discovered species and to hybrid strains originating from inter-species crosses.
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Mobile suitcase laboratory for rapid detection of Leishmania donovani using recombinase polymerase amplification assay.
Dinesh Mondal,Prakash Ghosh,Anik Ashfaq Khan,Faria Hossain,Susanne Böhlken-Fascher,Greg Matlashewski,Axel Kroeger,Piero Olliaro,Piero Olliaro,Ahmed Abd El Wahed +9 more
TL;DR: The mobile suitcase laboratory using RPA is ideal for rapid sensitive and specific detection of LD especially at low resource settings and could contribute to VL control and elimination programmes.
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Chagas Disease Diagnostic Applications: Present Knowledge and Future Steps.
TL;DR: In this article, the authors discuss the present knowledge and possible future steps in Chagas disease diagnostic applications, as well as the opportunity provided by recent advances in high-throughput methods for biomarker discovery.
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Loop-mediated isothermal amplification (LAMP): An advanced molecular point-of-care technique for the detection of Leishmania infection.
TL;DR: Observations suggest that LAMP offers a good molecular POC technique for the diagnosis of leishmaniasis and is also readily applicable to screening at-risk populations and vector sand flies for Leishmania infection in endemic areas.
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Journal ArticleDOI
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