{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,31]],"date-time":"2025-12-31T11:54:14Z","timestamp":1767182054429,"version":"build-2238731810"},"update-to":[{"DOI":"10.1371\/journal.pntd.0009530","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2021,8,5]],"date-time":"2021-08-05T00:00:00Z","timestamp":1628121600000}}],"reference-count":102,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2021,7,26]],"date-time":"2021-07-26T00:00:00Z","timestamp":1627257600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100012964","name":"Minist\u00e8re de l\u2019Enseignement Sup\u00e9rieur et de la Recherche Scientifique","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100012964","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Clinical Investigation Center, Institut Pasteur de Tunis"},{"name":"National Academy of Sciences-USAID-PEER Program","award":["AID-OAA-A-11-00012-PEER 518"],"award-info":[{"award-number":["AID-OAA-A-11-00012-PEER 518"]}]}],"content-domain":{"domain":["www.plosntds.org"],"crossmark-restriction":false},"short-container-title":["PLoS Negl Trop Dis"],"abstract":"\n Background<\/jats:title>\n \n Dipeptidyl peptidase III<\/jats:italic>\n (\n DPPIII<\/jats:italic>\n ) member of M49 peptidase family is a zinc-dependent metallopeptidase that cleaves dipeptides sequentially from the N-terminus of its substrates. In\n Leishmania<\/jats:italic>\n ,\n DPPIII<\/jats:italic>\n , was reported with other peptidases to play a significant role in parasites\u2019 growth and survival. In a previous study, we used a coding sequence annotated as\n DPPIII<\/jats:italic>\n to develop and evaluate a PCR assay that is specific to dermotropic Old World (OW)\n Leishmania<\/jats:italic>\n species. Thus, our objective was to further assess use of this gene for\n Leishmania<\/jats:italic>\n species identification and for phylogeny, and thus for diagnostic and molecular epidemiology studies of Old World\n Leishmania<\/jats:italic>\n species.\n <\/jats:p>\n <\/jats:sec>\n \n Methodology<\/jats:title>\n \n Orthologous\n DDPIII<\/jats:italic>\n genes were searched in all\n Leishmania<\/jats:italic>\n genomes and aligned to design PCR primers and identify relevant restriction enzymes. A PCR assays was developed and seventy-two\n Leishmania<\/jats:italic>\n fragment sequences were analyzed using MEGA X genetics software to infer evolution and phylogenetic relationships of studied species and strains. A PCR-RFLP scheme was also designed and tested on 58 OW\n Leishmania<\/jats:italic>\n strains belonging to 8\n Leishmania<\/jats:italic>\n species and evaluated on 75 human clinical skin samples.\n <\/jats:p>\n <\/jats:sec>\n \n Findings<\/jats:title>\n \n Sequence analysis showed 478 variable sites (302 being parsimony informative). Test of natural selection (dN-dS) (-0.164, SE = 0.013) inferred a negative selection, characteristic of essential genes, corroborating the\n DPPIII<\/jats:italic>\n importance for parasite survival. Inter- and intra-specific genetic diversity was used to develop universal amplification of a 662bp fragment. Sequence analyses and phylogenies confirmed occurrence of 6 clusters congruent to\n L<\/jats:italic>\n .\n major<\/jats:italic>\n ,\n L<\/jats:italic>\n .\n tropica<\/jats:italic>\n ,\n L<\/jats:italic>\n .\n aethiopica<\/jats:italic>\n ,\n L<\/jats:italic>\n .\n arabica<\/jats:italic>\n ,\n L<\/jats:italic>\n .\n turanica<\/jats:italic>\n ,\n L<\/jats:italic>\n .\n tarentolae<\/jats:italic>\n species, and one to the\n L<\/jats:italic>\n .\n infantum<\/jats:italic>\n and\n L<\/jats:italic>\n .\n donovani<\/jats:italic>\n species complex.\n <\/jats:p>\n \n A PCR-RFLP algorithm for\n Leishmania<\/jats:italic>\n species identification was designed using double digestions with\n Hae<\/jats:italic>\n III and\n Kpn<\/jats:italic>\n I and with\n Sac<\/jats:italic>\n I and\n Pvu<\/jats:italic>\n II endonucleases. Overall, this PCR-RFLP yielded distinct profiles for each of the species\n L<\/jats:italic>\n .\n major<\/jats:italic>\n ,\n L<\/jats:italic>\n .\n tropica<\/jats:italic>\n ,\n L<\/jats:italic>\n .\n aethiopica<\/jats:italic>\n ,\n L<\/jats:italic>\n .\n arabica<\/jats:italic>\n and\n L<\/jats:italic>\n .\n turanica<\/jats:italic>\n and the\n L<\/jats:italic>\n . (\n Sauroleishmania) L<\/jats:italic>\n .\n tarentolae<\/jats:italic>\n . The species\n L<\/jats:italic>\n .\n donovani<\/jats:italic>\n , and\n L<\/jats:italic>\n .\n infantum<\/jats:italic>\n shared the same profile except for strains of Indian origin. When tested on clinical samples, the\n DPPIII<\/jats:italic>\n PCR showed sensitivities of 82.22% when compared to direct examination and was able to identify 84.78% of the positive samples.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion<\/jats:title>\n \n The study demonstrates that\n DPPIII<\/jats:italic>\n gene is suitable to detect and identify\n Leishmania<\/jats:italic>\n species and to complement other molecular methods for leishmaniases diagnosis and epidemiology. Thus, it can contribute to evidence-based disease control and surveillance.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1371\/journal.pntd.0009530","type":"journal-article","created":{"date-parts":[[2021,7,26]],"date-time":"2021-07-26T13:38:57Z","timestamp":1627306737000},"page":"e0009530","update-policy":"https:\/\/doi.org\/10.1371\/journal.pntd.corrections_policy","source":"Crossref","is-referenced-by-count":5,"title":["Dipeptidyl peptidase III as a DNA marker to investigate epidemiology and taxonomy of Old World Leishmania species"],"prefix":"10.1371","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5289-5892","authenticated-orcid":true,"given":"Insaf","family":"Bel Hadj Ali","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5920-6113","authenticated-orcid":true,"given":"Hamed","family":"Chouaieb","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yusr","family":"Saadi Ben Aoun","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emna","family":"Harigua-Souiai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4455-5900","authenticated-orcid":true,"given":"Hejer","family":"Souguir","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alia","family":"Yaacoub","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oussa\u00efma","family":"El Dbouni","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zoubir","family":"Harrat","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2183-5915","authenticated-orcid":true,"given":"Maowia M.","family":"Mukhtar","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Moncef","family":"Ben Said","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nabil","family":"Haddad","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akila","family":"Fathallah-Mili","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ikram","family":"Guizani","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2021,7,26]]},"reference":[{"key":"pntd.0009530.ref001","first-page":"7","article-title":"Leishmaniasis Worldwide and Global Estimates of Its Incidence","author":"J Alvar","year":"2012","journal-title":"PLoS ONE"},{"key":"pntd.0009530.ref002","unstructured":"WHO. 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