A Simplified and Standardized Polymerase Chain Reaction Format for the Diagnosis of Leishmaniasis
Identifieur interne : 003241 ( Main/Exploration ); précédent : 003240; suivant : 003242A Simplified and Standardized Polymerase Chain Reaction Format for the Diagnosis of Leishmaniasis
Auteurs : Stijn Deborggraeve [Belgique] ; Thierry Laurent [Belgique] ; Diego Espinosa [Pérou] ; Gert Van Der Auwera [Belgique] ; Margaret Mbuchi [Kenya] ; Monique Wasunna [Kenya] ; Sayda El-Safi [Soudan] ; Ahmed Almustafa Al-Basheer [Soudan] ; Jorge Arévalo [Pérou] ; César Miranda-Verástegui [Pérou] ; Thierry Leclipteux [Belgique] ; Pascal Mertens [Belgique] ; Jean-Claude Dujardin [Belgique] ; Piet Herdewijn [Belgique] ; Philippe Büscher [Belgique]Source :
- The Journal of Infectious Diseases [ 0022-1899 ] ; 2008.
Abstract
Background. Definite diagnosis of Leishmania infections is based on demonstration of the parasite by microscopic analysis of tissue biopsy specimens or aspirate samples. However, microscopy generally shows low sensitivity and requires invasive sampling. Methods. We describe here the development of a simple and rapid test for the detection of polymerase chain reaction-amplified Leishmania DNA. A phase 1 evaluation of the text was conducted in clinical samples from 60 nonendemic and 45 endemic control subjects and from 44 patients with confirmed cutaneous leishmaniasis (CL), 12 with mucocutaneous leishmaniasis (MCL), and 43 with visceral leishmaniasis (VL) from Peru, Kenya, and Sudan. Results. The lower detection limits of the assay are 10 fg of Leishmania DNA and 1 parasite in 180 µL of blood. The specificity was 98.3% (95% confidence interval [CI], 91.1%–99.7%) and 95.6% (95% CI, 85.2%–98.8%) for nonendemic and endemic control samples, respectively, and the sensitivity was 93.2% (95% CI, 81.8%–97.7%), 91.7% (95% CI, 64.6%–98.5%), and 86% (95% CI, 72.7%–93.4%) for lesions from patients with CL or MCL and blood from patients with VL, respectively. Conclusions. The Leishmania OligoC-TesT showed high specificity and sensitivity in clinical samples and was able to detect the parasite in samples obtained by less invasive means, such as blood, lymph, and lesion scrapings. The assay is a promising new tool for simplified and standardized molecular detection of Leishmania parasites.
Url:
DOI: 10.1086/592509
Affiliations:
- Belgique, Kenya, Pérou, Soudan
- Province d'Anvers, Province du Brabant flamand, Région flamande
- Anvers, Louvain
- Katholieke Universiteit Leuven
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Antwerp</wicri:regionArea><wicri:noRegion>Antwerp</wicri:noRegion></affiliation><affiliation wicri:level="4"><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Medicinal Chemistry, Rega Institute for Medicinal Research, Katholieke Universiteit Leuven, Leuven</wicri:regionArea><orgName type="university">Katholieke Universiteit Leuven</orgName><placeName><settlement type="city">Louvain</settlement><region>Région flamande</region><region type="district" nuts="2">Province du Brabant flamand</region></placeName></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Belgique</country></affiliation><affiliation wicri:level="3"><country wicri:rule="url">Belgique</country><wicri:regionArea>Reprints or correspondence: Dr. Stijn Deborggraeve, Dept. of Parasitology, Institute of Tropical Medicine, Nationalestraat 155, 2000 Antwerp</wicri:regionArea><placeName><region type="province" nuts="2">Province d'Anvers</region><settlement type="city">Anvers</settlement></placeName></affiliation></author><author><name sortKey="Laurent, Thierry" sort="Laurent, Thierry" uniqKey="Laurent T" first="Thierry" last="Laurent">Thierry Laurent</name><affiliation wicri:level="1"><country xml:lang="fr">Belgique</country><wicri:regionArea>Coris BioConcept, Gembloux</wicri:regionArea><wicri:noRegion>Gembloux</wicri:noRegion></affiliation></author><author><name sortKey="Espinosa, Diego" sort="Espinosa, Diego" uniqKey="Espinosa D" first="Diego" last="Espinosa">Diego Espinosa</name><affiliation wicri:level="1"><country xml:lang="fr">Pérou</country><wicri:regionArea>Instituto de Medicina Tropical Alexander von Humboldt and Faculty of Sciences and Philosophy, Universidad Peruana Cayetano Heredia, Lima</wicri:regionArea><wicri:noRegion>Lima</wicri:noRegion></affiliation></author><author><name sortKey="Van Der Auwera, Gert" sort="Van Der Auwera, Gert" uniqKey="Van Der Auwera G" first="Gert" last="Van Der Auwera">Gert Van Der Auwera</name><affiliation wicri:level="1"><country xml:lang="fr">Belgique</country><wicri:regionArea>Department of Parasitology, Institute of Tropical Medicine Antwerp, Antwerp</wicri:regionArea><wicri:noRegion>Antwerp</wicri:noRegion></affiliation></author><author><name sortKey="Mbuchi, Margaret" sort="Mbuchi, Margaret" uniqKey="Mbuchi M" first="Margaret" last="Mbuchi">Margaret Mbuchi</name><affiliation wicri:level="1"><country xml:lang="fr">Kenya</country><wicri:regionArea>Center for Clinical Research, Kenya Medical Research Institute, Nairobi</wicri:regionArea><wicri:noRegion>Nairobi</wicri:noRegion></affiliation></author><author><name sortKey="Wasunna, Monique" sort="Wasunna, Monique" uniqKey="Wasunna M" first="Monique" last="Wasunna">Monique Wasunna</name><affiliation wicri:level="1"><country xml:lang="fr">Kenya</country><wicri:regionArea>Center for Clinical Research, Kenya Medical Research Institute, Nairobi</wicri:regionArea><wicri:noRegion>Nairobi</wicri:noRegion></affiliation></author><author><name sortKey="El Safi, Sayda" sort="El Safi, Sayda" uniqKey="El Safi S" first="Sayda" last="El-Safi">Sayda El-Safi</name><affiliation wicri:level="1"><country xml:lang="fr">Soudan</country><wicri:regionArea>Department of Microbiology and Parasitology, Khartoum University, Khartoum</wicri:regionArea><wicri:noRegion>Khartoum</wicri:noRegion></affiliation></author><author><name sortKey="Al Basheer, Ahmed Almustafa" sort="Al Basheer, Ahmed Almustafa" uniqKey="Al Basheer A" first="Ahmed Almustafa" last="Al-Basheer">Ahmed Almustafa Al-Basheer</name><affiliation wicri:level="1"><country xml:lang="fr">Soudan</country><wicri:regionArea>Department of Microbiology and Parasitology, Khartoum University, Khartoum</wicri:regionArea><wicri:noRegion>Khartoum</wicri:noRegion></affiliation></author><author><name sortKey="Arevalo, Jorge" sort="Arevalo, Jorge" uniqKey="Arevalo J" first="Jorge" last="Arévalo">Jorge Arévalo</name><affiliation wicri:level="1"><country xml:lang="fr">Pérou</country><wicri:regionArea>Instituto de Medicina Tropical Alexander von Humboldt and Faculty of Sciences and Philosophy, Universidad Peruana Cayetano Heredia, Lima</wicri:regionArea><wicri:noRegion>Lima</wicri:noRegion></affiliation></author><author><name sortKey="Miranda Verastegui, Cesar" sort="Miranda Verastegui, Cesar" uniqKey="Miranda Verastegui C" first="César" last="Miranda-Verástegui">César Miranda-Verástegui</name><affiliation wicri:level="1"><country xml:lang="fr">Pérou</country><wicri:regionArea>Instituto de Medicina Tropical Alexander von Humboldt and Faculty of Sciences and Philosophy, Universidad Peruana Cayetano Heredia, Lima</wicri:regionArea><wicri:noRegion>Lima</wicri:noRegion></affiliation></author><author><name sortKey="Leclipteux, Thierry" sort="Leclipteux, Thierry" uniqKey="Leclipteux T" first="Thierry" last="Leclipteux">Thierry Leclipteux</name><affiliation wicri:level="1"><country xml:lang="fr">Belgique</country><wicri:regionArea>Coris BioConcept, Gembloux</wicri:regionArea><wicri:noRegion>Gembloux</wicri:noRegion></affiliation></author><author><name sortKey="Mertens, Pascal" sort="Mertens, Pascal" uniqKey="Mertens P" first="Pascal" last="Mertens">Pascal Mertens</name><affiliation wicri:level="1"><country xml:lang="fr">Belgique</country><wicri:regionArea>Coris BioConcept, Gembloux</wicri:regionArea><wicri:noRegion>Gembloux</wicri:noRegion></affiliation></author><author><name sortKey="Dujardin, Jean Claude" sort="Dujardin, Jean Claude" uniqKey="Dujardin J" first="Jean-Claude" last="Dujardin">Jean-Claude Dujardin</name><affiliation wicri:level="1"><country xml:lang="fr">Belgique</country><wicri:regionArea>Department of Parasitology, Institute of Tropical Medicine Antwerp, Antwerp</wicri:regionArea><wicri:noRegion>Antwerp</wicri:noRegion></affiliation></author><author><name sortKey="Herdewijn, Piet" sort="Herdewijn, Piet" uniqKey="Herdewijn P" first="Piet" last="Herdewijn">Piet Herdewijn</name><affiliation wicri:level="4"><country xml:lang="fr">Belgique</country><wicri:regionArea>Laboratory of Medicinal Chemistry, Rega Institute for Medicinal Research, Katholieke Universiteit Leuven, Leuven</wicri:regionArea><orgName type="university">Katholieke Universiteit Leuven</orgName><placeName><settlement type="city">Louvain</settlement><region>Région flamande</region><region type="district" nuts="2">Province du Brabant flamand</region></placeName></affiliation></author><author><name sortKey="Buscher, Philippe" sort="Buscher, Philippe" uniqKey="Buscher P" first="Philippe" last="Büscher">Philippe Büscher</name><affiliation wicri:level="1"><country xml:lang="fr">Belgique</country><wicri:regionArea>Department of Parasitology, Institute of Tropical Medicine Antwerp, Antwerp</wicri:regionArea><wicri:noRegion>Antwerp</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">The Journal of Infectious Diseases</title><title level="j" type="abbrev">The Journal of Infectious Diseases</title><idno type="ISSN">0022-1899</idno><idno type="eISSN">1537-6613</idno><imprint><publisher>The University of Chicago Press</publisher><date type="published">2008</date><biblScope unit="vol">198</biblScope><biblScope unit="issue">10</biblScope><biblScope unit="page" from="1565">1565</biblScope><biblScope unit="page" to="1572">1572</biblScope></imprint><idno type="ISSN">0022-1899</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0022-1899</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract">Background. Definite diagnosis of Leishmania infections is based on demonstration of the parasite by microscopic analysis of tissue biopsy specimens or aspirate samples. However, microscopy generally shows low sensitivity and requires invasive sampling. Methods. We describe here the development of a simple and rapid test for the detection of polymerase chain reaction-amplified Leishmania DNA. A phase 1 evaluation of the text was conducted in clinical samples from 60 nonendemic and 45 endemic control subjects and from 44 patients with confirmed cutaneous leishmaniasis (CL), 12 with mucocutaneous leishmaniasis (MCL), and 43 with visceral leishmaniasis (VL) from Peru, Kenya, and Sudan. Results. The lower detection limits of the assay are 10 fg of Leishmania DNA and 1 parasite in 180 µL of blood. The specificity was 98.3% (95% confidence interval [CI], 91.1%–99.7%) and 95.6% (95% CI, 85.2%–98.8%) for nonendemic and endemic control samples, respectively, and the sensitivity was 93.2% (95% CI, 81.8%–97.7%), 91.7% (95% CI, 64.6%–98.5%), and 86% (95% CI, 72.7%–93.4%) for lesions from patients with CL or MCL and blood from patients with VL, respectively. Conclusions. The Leishmania OligoC-TesT showed high specificity and sensitivity in clinical samples and was able to detect the parasite in samples obtained by less invasive means, such as blood, lymph, and lesion scrapings. The assay is a promising new tool for simplified and standardized molecular detection of Leishmania parasites.</div></front></TEI><affiliations><list><country><li>Belgique</li><li>Kenya</li><li>Pérou</li><li>Soudan</li></country><region><li>Province d'Anvers</li><li>Province du Brabant flamand</li><li>Région flamande</li></region><settlement><li>Anvers</li><li>Louvain</li></settlement><orgName><li>Katholieke Universiteit Leuven</li></orgName></list><tree><country name="Belgique"><noRegion><name sortKey="Deborggraeve, Stijn" sort="Deborggraeve, Stijn" uniqKey="Deborggraeve S" first="Stijn" last="Deborggraeve">Stijn Deborggraeve</name></noRegion><name sortKey="Buscher, Philippe" sort="Buscher, Philippe" uniqKey="Buscher P" first="Philippe" last="Büscher">Philippe Büscher</name><name sortKey="Deborggraeve, Stijn" sort="Deborggraeve, Stijn" uniqKey="Deborggraeve S" first="Stijn" last="Deborggraeve">Stijn Deborggraeve</name><name sortKey="Deborggraeve, Stijn" sort="Deborggraeve, Stijn" uniqKey="Deborggraeve S" first="Stijn" last="Deborggraeve">Stijn Deborggraeve</name><name sortKey="Deborggraeve, Stijn" sort="Deborggraeve, Stijn" uniqKey="Deborggraeve S" first="Stijn" last="Deborggraeve">Stijn Deborggraeve</name><name sortKey="Dujardin, Jean Claude" sort="Dujardin, Jean Claude" uniqKey="Dujardin J" first="Jean-Claude" last="Dujardin">Jean-Claude Dujardin</name><name sortKey="Herdewijn, Piet" sort="Herdewijn, Piet" uniqKey="Herdewijn P" first="Piet" last="Herdewijn">Piet Herdewijn</name><name sortKey="Laurent, Thierry" sort="Laurent, Thierry" uniqKey="Laurent T" first="Thierry" last="Laurent">Thierry Laurent</name><name sortKey="Leclipteux, Thierry" sort="Leclipteux, Thierry" uniqKey="Leclipteux T" first="Thierry" last="Leclipteux">Thierry Leclipteux</name><name sortKey="Mertens, Pascal" sort="Mertens, Pascal" uniqKey="Mertens P" first="Pascal" last="Mertens">Pascal Mertens</name><name sortKey="Van Der Auwera, Gert" sort="Van Der Auwera, Gert" uniqKey="Van Der Auwera G" first="Gert" last="Van Der Auwera">Gert Van Der Auwera</name></country><country name="Pérou"><noRegion><name sortKey="Espinosa, Diego" sort="Espinosa, Diego" uniqKey="Espinosa D" first="Diego" last="Espinosa">Diego Espinosa</name></noRegion><name sortKey="Arevalo, Jorge" sort="Arevalo, Jorge" uniqKey="Arevalo J" first="Jorge" last="Arévalo">Jorge Arévalo</name><name sortKey="Miranda Verastegui, Cesar" sort="Miranda Verastegui, Cesar" uniqKey="Miranda Verastegui C" first="César" last="Miranda-Verástegui">César Miranda-Verástegui</name></country><country name="Kenya"><noRegion><name sortKey="Mbuchi, Margaret" sort="Mbuchi, Margaret" uniqKey="Mbuchi M" first="Margaret" last="Mbuchi">Margaret Mbuchi</name></noRegion><name sortKey="Wasunna, Monique" sort="Wasunna, Monique" uniqKey="Wasunna M" first="Monique" last="Wasunna">Monique Wasunna</name></country><country name="Soudan"><noRegion><name sortKey="El Safi, Sayda" sort="El Safi, Sayda" uniqKey="El Safi S" first="Sayda" last="El-Safi">Sayda El-Safi</name></noRegion><name sortKey="Al Basheer, Ahmed Almustafa" sort="Al Basheer, Ahmed Almustafa" uniqKey="Al Basheer A" first="Ahmed Almustafa" last="Al-Basheer">Ahmed Almustafa Al-Basheer</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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