A Simplified and Standardized Polymerase Chain Reaction Format for the Diagnosis of Leishmaniasis
| Autor: | Thierry Leclipteux, Pascal Mertens, Margaret Mbuchi, Thierry Laurent, Sayda El-Safi, Cesar Miranda-Verastegui, Monique Wasunna, Stijn Deborggraeve, Diego A. Espinosa, Philippe Büscher, Jorge Arevalo, Jean-Claude Dujardin, Piet Herdewijn, Ahmed Almustafa Al-Basheer, Gert Van der Auwera |
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| Rok vydání: | 2008 |
| Předmět: |
Performance
Molecular Sequence Data Polymerase Chain Reaction Sensitivity and Specificity Major Articles law.invention Major Articles and Brief Reports Sensitivity Cutaneous leishmaniasis law parasitic diseases Biopsy RNA Ribosomal 18S medicine Animals Humans Immunology and Allergy Parasite hosting Parasites Leishmaniasis Polymerase chain reaction Leishmania Base Sequence biology medicine.diagnostic_test Rapid diagnostic tests Protozoal diseases medicine.disease biology.organism_classification Virology PCR Infectious Diseases Visceral leishmaniasis Laboratory diagnosis Specificity Simplified Lymph Sequence Alignment Molecular diagnostic techniques |
| Zdroj: | The Journal of Infectious Diseases |
| ISSN: | 1537-6613 0022-1899 |
| DOI: | 10.1086/592509 |
| Popis: | 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 L 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 abletodetecttheparasiteinsamplesobtainedbylessinvasivemeans,suchasblood,lymph,andlesionscrapings.The assay is a promising new tool for simplified and standardized molecular detection of Leishmania parasites. Leishmaniasis is a vectorborne disease caused by protozoa of the genus Leishmania and is endemic in many areas of |
| Databáze: | OpenAIRE |
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