Gauging the skin resident Leishmania parasites through a loop mediated isothermal amplification (LAMP) assay in post-kala-azar dermal leishmaniasis

Ghosh, Prakash, Chowdhury, Rajashree, Maruf, Shomik, Picado, Albert, Hossain, Faria, Owen, Sophie ORCID: https://orcid.org/0000-0002-0458-2357, 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 ORCID: https://orcid.org/0000-0002-0816-2835, Ndungu, Joseph, Cruz, Israel and Mondal, Dinesh (2022) 'Gauging the skin resident Leishmania parasites through a loop mediated isothermal amplification (LAMP) assay in post-kala-azar dermal leishmaniasis'. Scientific Reports, Vol 12, Issue 1, e18069.

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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.

Item Type:	Article
Subjects:	QX Parasitology > Insects. Other Parasites > QX 505 Diptera WC Communicable Diseases > Tropical and Parasitic Diseases > WC 715 Visceral leishmaniasis WR Dermatology > Parasitic Skin Diseases > WR 345 Parasitic skin diseases (General or not elsewhere classified) WR Dermatology > Parasitic Skin Diseases > WR 350 Tropical diseases of the skin. Mucocutaneous leishmaniasis. Leishmaniasis
Faculty: Department:	Biological Sciences > Department of Tropical Disease Biology
Digital Object Identifer (DOI):	https://doi.org/10.1038/s41598-022-21497-6
SWORD Depositor:	JISC Pubrouter
Depositing User:	JISC Pubrouter
Date Deposited:	17 Nov 2022 14:03
Last Modified:	17 Nov 2022 14:03
URI:	https://archive.lstmed.ac.uk/id/eprint/21379

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