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 microL 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.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2008 |
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| Erschienen: |
2008 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:198 |
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| Enthalten in: |
The Journal of infectious diseases - 198(2008), 10 vom: 15. Nov., Seite 1565-72 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Deborggraeve, Stijn [VerfasserIn] |
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| Links: |
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| Themen: |
Journal Article |
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| Anmerkungen: |
Date Completed 06.01.2009 Date Revised 08.04.2022 published: Print Citation Status MEDLINE |
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| doi: |
10.1086/592509 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM182657833 |
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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 |
| 264 | 1 | |c 2008 | |
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| 500 | |a Date Revised 08.04.2022 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 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 | ||
| 520 | |a 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 | ||
| 520 | |a RESULTS: The lower detection limits of the assay are 10 fg of Leishmania DNA and 1 parasite in 180 microL 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 | ||
| 520 | |a 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 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a RNA, Ribosomal, 18S |2 NLM | |
| 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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