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{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,9,12]],"date-time":"2023-09-12T21:37:02Z","timestamp":1694554622705},"reference-count":42,"publisher":"Public Library of Science (PLoS)","issue":"4","license":[{"start":{"date-parts":[[2023,4,19]],"date-time":"2023-04-19T00:00:00Z","timestamp":1681862400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100008789","name":"Department of Science and Technology, Government of West Bengal","doi-asserted-by":"publisher","award":["969\/(Sanc)\/ST\/P\/SandT\/9G-22\/2016"]},{"DOI":"10.13039\/501100001411","name":"Indian Council of Medical Research","doi-asserted-by":"publisher","award":["6\/9-7(151)2017-ECD II"]},{"name":"JC Bose Fellowship, Govt. of India","award":["JCB\/2019\/000043"]},{"name":"Department Of Science & Technology, Govt. of India - German Academic Exchange Service","award":["DST\/INT\/DAAD\/P-13\/2020"]},{"name":"Department Of Science & Technology, Govt. of India - German Academic Exchange Service","award":["57588068"]}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"<jats:sec id=\"sec001\">\n<jats:title>Background<\/jats:title>\n<jats:p>The potential reservoirs of visceral leishmaniasis (VL) in South Asia include asymptomatic and relapsed cases of VL, along with patients with post kala-azar dermal leishmaniasis (PKDL). Accordingly, accurate estimation of their parasite load is pivotal for ensuring disease elimination, presently targeted for 2023. Serological tests cannot accurately detect relapses and\/or monitor treatment effectiveness, and therefore, parasite antigen\/nucleic acid based detection assays remain the only viable option. An excellent option is the quantitative polymerase chain reaction (qPCR) but the high cost, technical expertise and time involved precludes its wider acceptability. Accordingly, the recombinase polymerase amplification (RPA) assay operated in a mobile suitcase laboratory has emerged not simply as a diagnostic tool for leishmaniasis but also to monitor the disease burden.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec002\">\n<jats:title>Methodology\/Principal findings<\/jats:title>\n<jats:p>Using total genomic DNA isolated from peripheral blood of confirmed VL cases (n = 40) and lesional biopsies of PKDL cases (n = 64), the kinetoplast-DNA based qPCR and RPA assay was performed and parasite load expressed as Cycle threshold (Ct) and Time threshold (Tt) respectively. Using qPCR as the gold standard, the diagnostic specificity and sensitivity of RPA in na\u00efve cases of VL and PKDL was reiterated. To assess the prognostic potential of the RPA, samples were analyzed immediately at the end of treatment or \u22656 months following completion of treatment. In cases of VL, the RPA assay in terms of cure and detection of a relapse case showed 100% concordance with qPCR. In PKDL following completion of treatment, the overall detection concordance between RPA and qPCR was 92.7% (38\/41). At the end of treatment for PKDL, 7 cases remained qPCR positive, whereas RPA was positive in only 4\/7 cases, perhaps attributable to their low parasite load.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec003\">\n<jats:title>Conclusions\/Significance<\/jats:title>\n<jats:p>This study endorsed the potential of RPA to evolve as a field applicable, molecular tool for monitoring parasite load, possibly at a point of care level and is worthy of consideration in resource limited settings.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pntd.0011231","type":"journal-article","created":{"date-parts":[[2023,4,19]],"date-time":"2023-04-19T17:32:48Z","timestamp":1681925568000},"page":"e0011231","update-policy":"http:\/\/dx.doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["Evaluation of Recombinase Polymerase Amplification assay for monitoring parasite load in patients with kala-azar a
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