Rapid differentiation of Old World Leishmania species by LightCycler polymerase chain reaction and melting curve analysis.
A LightCycler real-time polymerase chain reaction (PCR) assay has been developed to detect and differentiate four of the main Leishmania species of the Old World. The assay is based on fluorescence melting curve analysis of PCR products generated from the minicircles of kinetoplast DNA. According to the melting temperature, which is a function of GC/AT ratio, length and nucleotide sequences of the amplified product, Leishmania major was differentiated from L. donovani and from L. tropica and L. infantum. Melting curves analysis offers a rapid alternative for identification of species in diagnostic or epidemiological studies of leishmaniasis or asymptomatic parasitism.
Nicolas
Luc
L
Institut Pasteur, Unité d'Immunophysiologie et Parasitisme Intracellulaire, 25 rue du Dr Roux, 75724 Paris Cedex 15, France. lnicolas@pasteur.fr
Milon
Geneviève
G
Prina
Eric
E
D000818:Animals; D052160:Benzothiazoles; D015342:DNA Probes; D018105:DNA, Kinetoplast / Q000032:analysis*; D003959:Diamines; D005456:Fluorescent Dyes; D006801:Humans; D007891:Leishmania / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D007896:Leishmaniasis / Q000469:parasitology*; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013696:Temperature; D013997:Time Factors
D052160:Benzothiazoles; D015342:DNA Probes; D018105:DNA, Kinetoplast; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I