02243nas a2200253   4500000000100000008004100001260001600042653002400058653001700082653001900099653003100118653001200149653003300161100002700194700001300221700001200234700001100246700001300257245010000270300001100370490000800381520158600389022001401975       2023                            d  bElsevier BV10aInfectious Diseases10aParasitology10aInsect Science10aVeterinary (miscellaneous)10aMiniPCR10aCutaneous Leishmaniasis (CL)1 aCastellanos-Gonzalez A1 aCossio A1 aJojoa J1 aMoen S1 aTravi BL00aMiniPCR as a portable equipment for the molecular diagnosis of american cutaneous leishmaniasis  a1069260 v2433 a<p>There is an urgent need to improve the diagnostic capacity of&nbsp;cutaneous leishmaniasis&nbsp;(CL) in rural health centers to improve the management of the disease in patients from remote regions where the infection is endemic. Microscopy of Giemsa-stained lesion smears is the standard-of-care diagnostic test in virtually all health centers, but its sensitivity is suboptimal (50–70%) and prone to&nbsp;false negative results. We evaluated the performance of a low-cost&nbsp;DNA extraction&nbsp;buffer (LAB) using a portable miniPCR™ equipment coupled with an inexpensive fluorescence viewer to detect Leishmania DNA with the naked eye or using a commercial photo app. Using ten-fold&nbsp;serial dilutions&nbsp;of Leishmania (V.) panamensis&nbsp;promastigotes&nbsp;the miniPCR-F test detected 10&nbsp;parasites&nbsp;per µL, which was comparable to real-time PCR. Utilization of DNA from retrospective clinical samples preserved at -80&nbsp;°C from Colombia (<em>n</em>&nbsp;=&nbsp;28) or lesion exudate preserved in filter papers from Peru (<em>n</em>&nbsp;=&nbsp;48) showed that the miniPCR-fluorescent test had a 100% sensitivity and &gt; 90% specificity compared to real-time PCR. This study demonstrated the utility of LAB&nbsp;DNA extraction&nbsp;method for direct amplification of&nbsp;<em>Leishmania</em>&nbsp;using the miniPCR and reading of P51 results with the naked eye or via digital reading with a photo app. These preliminary results indicated that the miniPCR-F test workflow could be amenable to implementation in resource-limited health centers.</p>
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