{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T18:17:53Z","timestamp":1786558673773,"version":"build-2736575974"},"reference-count":41,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2022,5,9]],"date-time":"2022-05-09T00:00:00Z","timestamp":1652054400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Promotion of Science (JSPS)","award":["JP17H01685"],"award-info":[{"award-number":["JP17H01685"]}]},{"name":"Promotion of Science (JSPS)","award":["JP15H05283"],"award-info":[{"award-number":["JP15H05283"]}]},{"name":"Promotion of Science (JSPS)","award":["2019"],"award-info":[{"award-number":["2019"]}]},{"name":"Promotion of Science (JSPS)","award":["2019"],"award-info":[{"award-number":["2019"]}]},{"name":"Faculty of Medicine, University of Ruhuna, Sri Lanka","award":["JP17H01685"],"award-info":[{"award-number":["JP17H01685"]}]},{"name":"Faculty of Medicine, University of Ruhuna, Sri Lanka","award":["JP15H05283"],"award-info":[{"award-number":["JP15H05283"]}]},{"name":"Faculty of Medicine, University of Ruhuna, Sri Lanka","award":["2019"],"award-info":[{"award-number":["2019"]}]},{"name":"Faculty of Medicine, University of Ruhuna, Sri Lanka","award":["2019"],"award-info":[{"award-number":["2019"]}]},{"name":"Galle Medical Association, Sri Lanka","award":["JP17H01685"],"award-info":[{"award-number":["JP17H01685"]}]},{"name":"Galle Medical Association, Sri Lanka","award":["JP15H05283"],"award-info":[{"award-number":["JP15H05283"]}]},{"name":"Galle Medical Association, Sri Lanka","award":["2019"],"award-info":[{"award-number":["2019"]}]},{"name":"Galle Medical Association, Sri Lanka","award":["2019"],"award-info":[{"award-number":["2019"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>The recent surge in cutaneous leishmaniasis (CL) in Sri Lanka has rendered clinical diagnosis difficult; thus, laboratory confirmation is indispensable. A modified (two novel inner primers to detect CL caused by Leishmania donovani) nested Internal Transcribed Spacer-1 (ITS1) PCR-Restriction Fragment Length Polymorphism (RFLP) method was developed and tested. The sensitivity of the modified nested PCR was tested using serial dilutions (103 to 10\u22122) of the DNA extract of a cultured L. donovani DD8 strain. Patients (n = 194) from Southern Sri Lanka were examined clinically, microscopically (Slit Skin Smear-SSS) and using the modified nested PCR. The modified nested PCR detected 2.55 fg of parasite DNA compared to ITS1 PCR (25 fg) and detected more cases than SSS (94.3% vs. 77.3%; p &lt; 0.01). The RFLP pattern was L. donovani in all cases. The modified nested PCR performed well in clinically doubtful lesions (95% by PCR vs. 60% by SSS; p &lt; 0.01), ulcerated nodules (91% vs. 71.8%; p &lt; 0.01) and plaques (100% vs. 66.7%; p &lt; 0.01). SSS demonstrated sensitivity (80.9%), specificity (81.8%), PPV (98.7%) and NPV (20.5%) against modified PCR. Low parasite loads and atypical lesions can be diagnosed by the proposed method with higher accuracy.<\/jats:p>","DOI":"10.3390\/microorganisms10050990","type":"journal-article","created":{"date-parts":[[2022,5,10]],"date-time":"2022-05-10T00:30:28Z","timestamp":1652142628000},"page":"990","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Development of a Highly Sensitive Nested PCR and Its Application for the Diagnosis of Cutaneous Leishmaniasis in Sri Lanka"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8895-5415","authenticated-orcid":false,"given":"Nirmitha Lalindi","family":"De Silva","sequence":"first","affiliation":[{"name":"Department of Parasitology, Faculty of Medicine, University of Ruhuna, Galle 80000, Sri Lanka"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Viraji Nefertiti Hiromel","family":"De Silva","sequence":"additional","affiliation":[{"name":"Base Hospital Tangalle, Tangalle 82200, Sri Lanka"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arachchige Theja Hemapala","family":"Deerasinghe","sequence":"additional","affiliation":[{"name":"District General Hospital Hambantota, Hambantota 82000, Sri Lanka"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6259-616X","authenticated-orcid":false,"given":"Upeksha Lakmini","family":"Rathnapala","sequence":"additional","affiliation":[{"name":"School of Biosciences, University of Melbourne, Melbourne, VIC 3010, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Makoto","family":"Itoh","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Aichi Medical University School of Medicine, Aichi 480-1195, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hidekazu","family":"Takagi","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Aichi Medical University School of Medicine, Aichi 480-1195, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mirani Vasanthamala","family":"Weerasooriya","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Faculty of Medicine, University of Ruhuna, Galle 80000, Sri Lanka"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5429-9536","authenticated-orcid":false,"given":"Hirotomo","family":"Kato","sequence":"additional","affiliation":[{"name":"Division of Medical Zoology, Department of Infection and Immunity, Jichi Medical University, Tochigi 329-0498, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4981-8914","authenticated-orcid":false,"given":"Thishan Channa","family":"Yahathugoda","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Faculty of Medicine, University of Ruhuna, Galle 80000, Sri Lanka"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,5,9]]},"reference":[{"key":"ref_1","unstructured":"World Health Organization (2020, August 06). 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