A Simplified and Standardized Polymerase Chain Reaction Format for the Diagnosis of Leishmaniasis
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 ¼L 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..
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E-Artikel |
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Erscheinungsjahr: |
2008 |
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Erschienen: |
2008 |
Enthalten in: |
Zur Gesamtaufnahme - volume:198 |
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Sprache: |
Englisch |
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Beteiligte Personen: |
Deborggraeve, Stijn [VerfasserIn] |
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Themen: |
Applied sciences |
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PPN (Katalog-ID): |
JST048956244 |
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100 | 1 | |a Deborggraeve, Stijn |e verfasserin |4 aut | |
245 | 1 | 2 | |a A Simplified and Standardized Polymerase Chain Reaction Format for the Diagnosis of Leishmaniasis |
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520 | |a 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 ¼L 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. | ||
540 | |a Copyright 2008 Infectious Diseases Society of America | ||
650 | 4 | |a Applied sciences |x Laboratory techniques |x Nucleic acid amplification techniques |x Polymerase chain reaction | |
650 | 4 | |a Biological sciences |x Biology |x Physiology |x Body composition |x Body fluids |x Blood | |
650 | 4 | |a Biological sciences |x Ecology |x Ecological processes |x Ecosystem dynamics |x Trophic dynamics |x Trophic levels |x Heterotrophs |x Parasites | |
650 | 4 | |a Health sciences |x Medical conditions |x Diseases |x Skin diseases |x Infectious skin diseases |x Parasitic skin diseases |x Leishmaniasis | |
650 | 4 | |a Health sciences |x Medical conditions |x Physical trauma |x Lesions | |
650 | 4 | |a Health sciences |x Medical conditions |x Infections | |
650 | 4 | |a Health sciences |x Medical diagnosis |x Diagnostic methods |x Cytodiagnosis |x Biopsies | |
650 | 4 | |a Health sciences |x Medical conditions |x Diseases |x Skin diseases |x Infectious skin diseases |x Parasitic skin diseases |x Leishmaniasis |x Visceral leishmaniasis | |
650 | 4 | |a Physical sciences |x Chemistry |x Chemical compounds |x Chemicals |x Acids |x Nucleic acids |x DNA | |
650 | 4 | |a Physical sciences |x Earth sciences |x Geology |x Mineralogy |x Specimens | |
650 | 4 | |a Applied sciences |x Laboratory techniques |x Nucleic acid amplification techniques |x Polymerase chain reaction | |
650 | 4 | |a Biological sciences |x Biology |x Physiology |x Body composition |x Body fluids |x Blood | |
650 | 4 | |a Biological sciences |x Ecology |x Ecological processes |x Ecosystem dynamics |x Trophic dynamics |x Trophic levels |x Heterotrophs |x Parasites | |
650 | 4 | |a Health sciences |x Medical conditions |x Diseases |x Skin diseases |x Infectious skin diseases |x Parasitic skin diseases |x Leishmaniasis | |
650 | 4 | |a Health sciences |x Medical conditions |x Physical trauma |x Lesions | |
650 | 4 | |a Health sciences |x Medical conditions |x Infections | |
650 | 4 | |a Health sciences |x Medical diagnosis |x Diagnostic methods |x Cytodiagnosis |x Biopsies | |
650 | 4 | |a Health sciences |x Medical conditions |x Diseases |x Skin diseases |x Infectious skin diseases |x Parasitic skin diseases |x Leishmaniasis |x Visceral leishmaniasis | |
650 | 4 | |a Physical sciences |x Chemistry |x Chemical compounds |x Chemicals |x Acids |x Nucleic acids |x DNA | |
650 | 4 | |a Physical sciences |x Earth sciences |x Geology |x Mineralogy |x Specimens |x Major Articles and Brief Reports |x Parasites | |
655 | 4 | |a research-article | |
700 | 1 | |a Laurent, Thierry |e verfasserin |4 aut | |
700 | 1 | |a Espinosa, Diego |e verfasserin |4 aut | |
700 | 1 | |a Van der Auwera, Gert |e verfasserin |4 aut | |
700 | 1 | |a Mbuchi, Margaret |e verfasserin |4 aut | |
700 | 1 | |a Wasunna, Monique |e verfasserin |4 aut | |
700 | 1 | |a El-Safi, Sayda |e verfasserin |4 aut | |
700 | 1 | |a Al-Basheer, Ahmed Almustafa |e verfasserin |4 aut | |
700 | 1 | |a Arévalo, Jorge |e verfasserin |4 aut | |
700 | 1 | |a Miranda-Verástegui, César |e verfasserin |4 aut | |
700 | 1 | |a Leclipteux, Thierry |e verfasserin |4 aut | |
700 | 1 | |a Mertens, Pascal |e verfasserin |4 aut | |
700 | 1 | |a Dujardin, Jean-Claude |e verfasserin |4 aut | |
700 | 1 | |a Herdewijn, Piet |e verfasserin |4 aut | |
700 | 1 | |a Büscher, Philippe |e verfasserin |4 aut | |
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