{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,2]],"date-time":"2026-03-02T21:22:56Z","timestamp":1772486576426,"version":"3.50.1"},"reference-count":41,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2003,4,1]],"date-time":"2003-04-01T00:00:00Z","timestamp":1049155200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2003,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Real-time technology eliminates many of the pitfalls of diagnostic PCR, but this method has not been applied to differentiation of\n            <jats:italic>Leishmania<\/jats:italic>\n            organisms so far. We have developed a real-time PCR that simultaneously detects, quantitates, and categorizes\n            <jats:italic>Leishmania<\/jats:italic>\n            organisms into three relevant groups causing distinct clinical pictures. The analytical sensitivity (detection rate of \u226595% at 94.1 parasites\/ml of blood) was within a range that has been determined previously to facilitate the confirmation of visceral leishmaniasis from peripheral blood. Parasites were successfully detected in 12 different clinical samples (blood, bone marrow, skin, and liver). The\n            <jats:italic>Leishmania donovani<\/jats:italic>\n            complex, the\n            <jats:italic>Leishmania brasiliensis<\/jats:italic>\n            complex, and species other than these could be clearly discriminated by means of distinct melting temperatures obtained with fluorescence resonance energy transfer probes (melting points, 72.7, 67.1, and 65.0\u00b0C, respectively). All three groups could be quantified within equal ranges. As in other real-time PCRs, the variability in the quantification of DNA was small (coefficient of variation [CV], &lt;2%). However, human samples containing low levels of parasites (100 parasites per ml of blood) showed higher variation (CV, 60.89%). Therefore, despite its superior analytical performance, care must be taken when real-time PCR is utilized for therapy monitoring.\n          <\/jats:p>","DOI":"10.1128\/jcm.41.4.1529-1535.2003","type":"journal-article","created":{"date-parts":[[2003,4,7]],"date-time":"2003-04-07T20:34:54Z","timestamp":1049747694000},"page":"1529-1535","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":93,"title":["Detection, Differentiation, and Quantitation of Pathogenic\n            <i>Leishmania<\/i>\n            Organisms by a Fluorescence Resonance Energy Transfer-Based Real-Time PCR Assay"],"prefix":"10.1128","volume":"41","author":[{"given":"Alexandra","family":"Schulz","sequence":"first","affiliation":[{"name":"National Reference Center for Tropical Infections"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Katja","family":"Mellenthin","sequence":"additional","affiliation":[{"name":"Leishmaniasis Group, Bernhard Nocht Institute for Tropical Medicine, Hamburg"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gabriele","family":"Scho\u0308nian","sequence":"additional","affiliation":[{"name":"Institute of Microbiology and Hygiene, Humboldt University, Charite\u0301, Berlin, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bernhard","family":"Fleischer","sequence":"additional","affiliation":[{"name":"National Reference Center for Tropical Infections"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christian","family":"Drosten","sequence":"additional","affiliation":[{"name":"National Reference Center for Tropical Infections"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.10.2.298"},{"key":"e_1_3_2_3_2","unstructured":"Anonymous. 2000. The leishmaniases and Leishmania \/HIV co-infections. WHO fact sheet 116. World Health Organization Geneva Switzerland."},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.1986.35.732"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/CDLI.8.4.828-831.2001"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-83-8-1887"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.2165\/00128071-200203020-00003"},{"key":"e_1_3_2_8_2","unstructured":"Desjeux P. J. P. Meert B. Piot J. Alvar F. J. Medrano M. Portus C. Munoz F. Laguna R. Lopez Velez A. Salas G. Sirera R. Cisterna C. Montalban H. Quero L. Gradoni M. Gramiccia R. Russo J. P. Dedet F. Pratlong J. Dereure M. Deniau A. Izri S. Matheron F. Farault P. Marty E. Rosenthal F. Antunes P. Abranche and R. Pradinaud. 2000. Leishmania\/HIV co-infection in south-western Europe 1990-1998: retrospective analysis of 965 cases. WHO\/Leish\/2000.40. 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