Please use this identifier to cite or link to this item:http://hdl.handle.net/20.500.12105/15944
Title
Gauging the skin resident Leishmania parasites through a loop mediated isothermal amplification (LAMP) assay in post-kala-azar dermal leishmaniasis
Author(s)
Ghosh, Prakash | Chowdhury, Rajashree | Maruf, Shomik | Picado, Albert | Hossain, Faria | Owen, Sophie I | Nath, Rupen | Baker, James | Hasnain, Md Golam | Shomik, Mohammad Sohel | Ghosh, Debashis | Rashid, Masud | Rashid, Md Utba | Sagar, Soumik Kha | Rahat, Md Abu | Basher, Ariful | Nath, Proggananda | Edwards, Thomas | Andrews, Jason R | Duthie, Malcolm S | de Souza, Dziedzom K | Adams, Emily R | Ndungu, Joseph | Cruz, Israel ISCIII | Mondal, Dinesh
Date issued
2022-10-27
Citation
Sci Rep. 2022 Oct 27;12(1):18069.
Language
Inglés
Document type
journal article
Abstract
Despite the availability of highly sensitive polymerase chain reaction (PCR)-based methods, the dearth of remotely deployable diagnostic tools circumvents the early and accurate detection of individuals with post-kala-azar dermal leishmaniasis (PKDL). Here, we evaluate a design-locked loop-mediated isothermal amplification (LAMP) assay to diagnose PKDL. A total of 76 snip-skin samples collected from individuals with probable PKDL (clinical presentation and a positive rK39 rapid diagnostic test (RDT)) were assessed by microscopy, qPCR, and LAMP. An equal number of age and sex-matched healthy controls were included to determine the specificity of the LAMP assay. The LAMP assay with a Qiagen DNA extraction (Q-LAMP) showed a promising sensitivity of 72.37% (95% CI: 60.91-82.01%) for identifying the PKDL cases. LAMP assay sensitivity declined when the DNA was extracted using a boil-spin method. Q-qPCR showed 68.42% (56.75-78.61%) sensitivity, comparable to LAMP and with an excellent agreement, whereas the microscopy exhibited a weak sensitivity of 39.47% (28.44-51.35%). When microscopy and/or qPCR were considered the gold standard, Q-LAMP exhibited an elevated sensitivity of 89.7% (95% CI: 78.83-96.11%) for detection of PKDL cases and Bayesian latent class modeling substantiated the excellent sensitivity of the assay. All healthy controls were found to be negative. Notwithstanding the optimum efficiency of the LAMP assay towards the detection of PKDL cases, further optimization of the boil-spin method is warranted to permit remote use of the assay.
MESH
Leishmaniasis, Visceral | Leishmania donovani | Parasites | Leishmaniasis, Cutaneous | Skin Diseases, Parasitic | Animals | Humans | Bayes Theorem | Real-Time Polymerase Chain Reaction
Online version
DOI
Collections