title: A new PCR-ELISA for diagnosis of visceral leishmaniasis in blood of HIV-negative subjects.
abstract: The PCR-ELISA represents a promising advance for diagnosis of visceral leishmaniasis (VL) in blood samples. However, the method has been validated mostly with HIV-positive patients who are known to have high levels of parasitaemia. We developed a new PCR-ELISA assay for specific detection of Leishmania in patients' blood and validated it in Nepalese subjects with clinically suspected VL, almost all of whom were HIV-negative. For blood samples, PCR-ELISA was more sensitive (83.9%) than conventional PCR (73.2%), and demonstrated 100% and 87.2% specificity when using healthy controls who had never travelled to a VL-endemic area and controls from a VL-endemic area as references, respectively. We have demonstrated the ability of PCR-ELISA to detect parasites in blood of HIV-negative patients. The method could be used for epidemiological as well as clinical purposes, as it reduces the need for traumatic bone marrow sampling and risky spleen aspiration.
authors.affiliation: Laboratory of Molecular Parasitology, Prins Leopold Instituut voor Tropische Geneeskunde, Nationalestraat 155, B-2000 Antwerp, Belgium.
authors.forename: Simonne
authors.initials: S
authors.lastname: De Doncker
authors.forename: Veronik
authors.initials: V
authors.lastname: Hutse
authors.forename: Saïd
authors.initials: S
authors.lastname: Abdellati
authors.forename: Suman
authors.initials: S
authors.lastname: Rijal
authors.forename: Bal Man
authors.initials: BM
authors.lastname: Singh Karki
authors.forename: Saskia
authors.initials: S
authors.lastname: Decuypere
authors.forename: Diane
authors.initials: D
authors.lastname: Jacquet
authors.forename: Dominique
authors.initials: D
authors.lastname: Le Ray
authors.forename: Marleen
authors.initials: M
authors.lastname: Boelaert
authors.forename: Shekhar
authors.initials: S
authors.lastname: Koirala
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Dujardin
mesh_terms: D000818:Animals; D016054:DNA, Protozoan / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D019353:Endemic Diseases / Q000517:prevention & control; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D018023:HIV Seronegativity / Q000502:physiology; D006801:Humans; D007893:Leishmania donovani / Q000235:genetics / Q000302:isolation & purification; D007898:Leishmaniasis, Visceral / Q000175:diagnosis* / Q000453:epidemiology / Q000469:parasitology; D009390:Nepal / Q000453:epidemiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D016054:DNA, Protozoan; D004275:DNA, Ribosomal