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.
Deborggraeve
Stijn
S
Department of Parasitology, Institute of Tropical Medicine Antwerp, Antwerp, Belgium. sdeborggraeve@itg.be
Laurent
Thierry
T
Espinosa
Diego
D
Van der Auwera
Gert
G
Mbuchi
Margaret
M
Wasunna
Monique
M
El-Safi
Sayda
S
Al-Basheer
Ahmed Almustafa
AA
Arévalo
Jorge
J
Miranda-Verástegui
César
C
Leclipteux
Thierry
T
Mertens
Pascal
P
Dujardin
Jean-Claude
JC
Herdewijn
Piet
P
Büscher
Philippe
P
D000818:Animals; D001483:Base Sequence; D006801:Humans; D007891:Leishmania / Q000235:genetics* / Q000302:isolation & purification; D007896:Leishmaniasis / Q000097:blood / Q000175:diagnosis*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012680:Sensitivity and Specificity; D016415:Sequence Alignment
D012337:RNA, Ribosomal, 18S