02917nas a2200337   4500000000100000008004100001260004400042653002900086653002400115653002600139653000900165100001100174700001400185700001300199700002200212700001200234700001300246700001100259700001200270700001200282700001300294700002000307700001700327700001300344245010800357856009300465300000800558490000700566520199200573022001402565       2024                            d  bSpringer Science and Business Media LLC10aLeishmaniasis, Cutaneous10aMolecular diagnosis10aLeishmania aethiopica10aLAMP1 aTaye B1 aMelkamu R1 aTajebe F1 aIbarra-Meneses AV1 aAdane D1 aAtnafu S1 aAdem M1 aAdane G1 aKassa M1 aAsres MS1 avan Griensven J1 avan Henten S1 aPareyn M00aEvaluation of Loopamp Leishmania detection kit for the diagnosis of cutaneous leishmaniasis in Ethiopia  uhttps://parasitesandvectors.biomedcentral.com/counter/pdf/10.1186/s13071-024-06475-3.pdf  a1-80 v173 a<p><strong>Background:</strong>&nbsp;Cutaneous leishmaniasis (CL) in Ethiopia and some parts of Kenya is predominantly caused by Leishmania aethiopica. While skin-slit (SS) microscopy is routinely used for CL diagnosis, more sensitive molecular tests are available. The Loopamp™ Leishmania detection kit (Loopamp) is a robust loop-mediated isothermal amplification (LAMP) assay with the potential for implementation in primary healthcare facilities. In this study, we comparatively assessed the diagnostic accuracy of four methods currently used to diagnose CL: Loopamp, kinetoplast DNA (kDNA) PCR, spliced leader RNA (SL-RNA) PCR and SS microscopy.</p>

<p><strong>Methods: </strong>A study on 122 stored tape disc samples of suspected CL patients was conducted in Gondar, northwestern Ethiopia. Routine SS microscopy results were obtained from all patients. Total nucleic acids were extracted from the tapes and subjected to PCR testing targeting kDNA and SL-RNA, and Loopamp. Diagnostic accuracy was calculated with SS microscopy as a reference test. The limit of detection (LoD) of Loopamp and kDNA PCR were determined for cultured L. aethiopica and Leishmania donovani.</p>

<p><strong>Results: </strong>Of the 122 patients, 64 (52.5%) were identified as CL cases based on SS microscopy. Although the PCR tests showed a sensitivity of 95.3% (95% confidence interval [CI] 91.6–99.1), Loopamp only had 48.4% (95% CI 39.6–57.3) sensitivity and 87.9% (95% CI 82.1–93.7) specificity. The LoD of Loopamp for L. donovani was 100-fold lower (20&nbsp;fg/µl) than that for L. aethiopica (2&nbsp;pg/µl).</p>

<p><strong>Conclusions: </strong>The Loopamp™ Leishmania detection kit is not suitable for the diagnosis of CL in Ethiopia, presumably due to a primer mismatch with the L. aethiopica 18S rRNA target. Further research is needed to develop a simple and sensitive point-of-care test that allows the decentralization of CL diagnosis in Ethiopia. Graphical Abstract</p>
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