Rapid detection of point mutations in Plasmodium falciparum genes associated with antimalarial drugs resistance by using High-Resolution Melting analysis.
We have developed a High-Resolution DNA Melting method to detect mutations related to Plasmodium falciparum resistance. This method is based on real-time PCR followed by High Resolution Melting ramping from 67 degrees C to 80 degrees C with fluorescence data acquisition set at 0.1 degrees C increments. The accuracy of the technique was assessed using 177 P. falciparum clinical isolates and two reference strains. Results perfectly matched those obtained by DNA sequencing for some important genetic markers of P. falciparum resistance. This technique could be of great value for epidemiological studies, especially in developing countries.
Andriantsoanirina
Valérie
V
Malaria Unit Research, Institut Pasteur de Madagascar, Antananarivo, Madagascar.
Lascombes
Vincent
V
Ratsimbasoa
Arsène
A
Bouchier
Christiane
C
Hoffman
Jonathan
J
Tichit
Magali
M
Rabarijaona
Leon-Paul
LP
Durand
Rémy
R
Ménard
Didier
D
D000818:Animals; D000962:Antimalarials / Q000494:pharmacology*; D016054:DNA, Protozoan / Q000235:genetics*; D004351:Drug Resistance; D006358:Hot Temperature; D006801:Humans; D009691:Nucleic Acid Denaturation; D021261:Parasitic Sensitivity Tests / Q000379:methods*; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000235:genetics; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D044366:Transition Temperature
D000962:Antimalarials; D016054:DNA, Protozoan