{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T16:23:09Z","timestamp":1792513389457,"version":"4.5.0"},"reference-count":40,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2021,3,12]],"date-time":"2021-03-12T00:00:00Z","timestamp":1615507200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000865","name":"Bill and Melinda Gates Foundation","doi-asserted-by":"publisher","award":["OPP53460"],"award-info":[{"award-number":["OPP53460"]}],"id":[{"id":"10.13039\/100000865","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>With the advancement of isothermal nucleic acid amplification techniques, detection of the pathogenic DNA in clinical samples at point-of-need is no longer a dream. The newly developed recombinase polymerase amplification (RPA) assay incorporated in a suitcase laboratory has shown promising diagnostic efficacy over real-time PCR in detection of leishmania DNA from clinical samples. For broader application of this point-of-need system, we undertook a current multi-country diagnostic evaluation study towards establishing this technique in different endemic settings which would be beneficial for the ongoing elimination programs for leishmaniasis. For this study purpose, clinical samples from confirmed visceral leishmaniasis (VL) and post-kala-azar dermal leishmaniasis (PKDL) patients were subjected to both real-time PCR and RPA assay in Bangladesh, India, and Nepal. Further skin samples from confirmed cutaneous leishmaniasis (CL) patients were also included from Sri Lanka. A total of 450 clinical samples from VL patients, 429 from PKDL patients, 47 from CL patients, and 322 from endemic healthy\/healthy controls were under investigation to determine the diagnostic efficacy of RPA assay in comparison to real-time PCR. A comparative sensitivity of both methods was found where real-time PCR and RPA assay showed 96.86% (95% CI: 94.45\u201398.42) and 88.85% (95% CI: 85.08\u201391.96) sensitivity respectively in the diagnosis of VL cases. This new isothermal method also exhibited promising diagnostic sensitivity (93.50%) for PKDL cases, when a skin sample was used. Due to variation in the sequence of target amplicons, RPA assay showed comparatively lower sensitivity (55.32%) than that of real-time PCR in Sri Lanka for the diagnosis of CL cases. Except for India, the assay presented absolute specificity in the rest of the sites. Excellent concordance between the two molecular methods towards detection of leishmania DNA in clinical samples substantiates the application of RPA assay incorporated in a suitcase laboratory for point-of-need diagnosis of VL and PKDL in low resource endemic settings. However, further improvisation of the method is necessary for diagnosis of CL.<\/jats:p>","DOI":"10.3390\/microorganisms9030588","type":"journal-article","created":{"date-parts":[[2021,3,14]],"date-time":"2021-03-14T23:52:06Z","timestamp":1615765926000},"page":"588","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["A Multi-Country, Single-Blinded, Phase 2 Study to Evaluate a Point-of-Need System for Rapid Detection of Leishmaniasis and Its Implementation in Endemic Settings"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4300-8761","authenticated-orcid":false,"given":"Prakash","family":"Ghosh","sequence":"first","affiliation":[{"name":"Nutrition and Clinical Service Division, International Centre for Diarrhoeal Disease Research, Dhaka 1212, Bangladesh"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abhijit","family":"Sharma","sequence":"additional","affiliation":[{"name":"PATH, 15th Floor, Dr. Gopal Das Bhawan, 28 Barakhamba Road, New Delhi 110001, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Narayan Raj","family":"Bhattarai","sequence":"additional","affiliation":[{"name":"B. P. Koirala Institute of Health Sciences, Dharan 56700, Nepal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kumar","family":"Abhishek","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, ICMR-Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna 800007, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2194-6802","authenticated-orcid":false,"given":"Thilini","family":"Nisansala","sequence":"additional","affiliation":[{"name":"Department of Microbiology, University of Sri Jayewardenepura, Gangodawila, Nugegoda 10250, Sri Lanka"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amresh","family":"Kumar","sequence":"additional","affiliation":[{"name":"PATH, 15th Floor, Dr. Gopal Das Bhawan, 28 Barakhamba Road, New Delhi 110001, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susanne","family":"B\u00f6hlken-Fascher","sequence":"additional","affiliation":[{"name":"Department of Animal Science, Division of Microbiology and Animal Hygiene, Georg-August-University of Goettingen, D-37077 Goettingen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rajashree","family":"Chowdhury","sequence":"additional","affiliation":[{"name":"Nutrition and Clinical Service Division, International Centre for Diarrhoeal Disease Research, Dhaka 1212, Bangladesh"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md Anik Ashfaq","family":"Khan","sequence":"additional","affiliation":[{"name":"Nutrition and Clinical Service Division, International Centre for Diarrhoeal Disease Research, Dhaka 1212, Bangladesh"},{"name":"Faculty of Medicine, University of Leipzig, D-04103 Leipzig, Germany"},{"name":"Institute of Animal Hygiene and Veterinary Public Health, University of Leipzig, D-04103 Leipzig, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Khaledul","family":"Faisal","sequence":"additional","affiliation":[{"name":"Nutrition and Clinical Service Division, International Centre for Diarrhoeal Disease Research, Dhaka 1212, Bangladesh"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Faria","family":"Hossain","sequence":"additional","affiliation":[{"name":"Nutrition and Clinical Service Division, International Centre for Diarrhoeal Disease Research, Dhaka 1212, Bangladesh"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Md. Rasel","family":"Uddin","sequence":"additional","affiliation":[{"name":"Nutrition and Clinical Service Division, International Centre for Diarrhoeal Disease Research, Dhaka 1212, Bangladesh"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2530-9982","authenticated-orcid":false,"given":"Md. Utba","family":"Rashid","sequence":"additional","affiliation":[{"name":"Nutrition and Clinical Service Division, International Centre for Diarrhoeal Disease Research, Dhaka 1212, Bangladesh"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8463-5326","authenticated-orcid":false,"given":"Shomik","family":"Maruf","sequence":"additional","affiliation":[{"name":"Nutrition and Clinical Service Division, International Centre for Diarrhoeal Disease Research, Dhaka 1212, Bangladesh"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9747-3431","authenticated-orcid":false,"given":"Keshav","family":"Rai","sequence":"additional","affiliation":[{"name":"B. P. Koirala Institute of Health Sciences, Dharan 56700, Nepal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monica","family":"Sooriyaarachchi","sequence":"additional","affiliation":[{"name":"Department of Parasitology, University of Sri Jayewardenepura, Gangodawila, Nugegoda 10250, Sri Lanka"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Withanage Lakma Kumari","family":"Abhayarathna","sequence":"additional","affiliation":[{"name":"Department of Parasitology, University of Sri Jayewardenepura, Gangodawila, Nugegoda 10250, Sri Lanka"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Prahlad","family":"Karki","sequence":"additional","affiliation":[{"name":"B. P. Koirala Institute of Health Sciences, Dharan 56700, Nepal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shiril","family":"Kumar","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, ICMR-Rajendra Memorial Research Institute of Medical Sciences, Agamkuan, Patna 800007, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shalindra","family":"Ranasinghe","sequence":"additional","affiliation":[{"name":"Department of Parasitology, University of Sri Jayewardenepura, Gangodawila, Nugegoda 10250, Sri Lanka"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Basudha","family":"Khanal","sequence":"additional","affiliation":[{"name":"B. P. 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