A new PCR-ELISA for diagnosis of visceral leishmaniasis in blood of HIV-negative subjects.
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.
De Doncker
Simonne
S
Laboratory of Molecular Parasitology, Prins Leopold Instituut voor Tropische Geneeskunde, Nationalestraat 155, B-2000 Antwerp, Belgium.
Hutse
Veronik
V
Abdellati
Saïd
S
Rijal
Suman
S
Singh Karki
Bal Man
BM
Decuypere
Saskia
S
Jacquet
Diane
D
Le Ray
Dominique
D
Boelaert
Marleen
M
Koirala
Shekhar
S
Dujardin
Jean-Claude
JC
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
D016054:DNA, Protozoan; D004275:DNA, Ribosomal