A new method for detection of pfmdr1 mutations in Plasmodium falciparum DNA using real-time PCR.
BACKGROUND
Surveillance for drug-resistant Plasmodium falciparum should be a component of malaria control programmes. Real-time PCR methods for the detection of parasite single-nucleotide polymorphisms (SNPs) and gene amplification could be useful survellance tools.


METHODS
A real-time PCR assay has been developed that identifies single nucleotide polymorphisms (SNPs) at amino acids 86, 184, 1034 and 1042 in the P. falciparum multi-drug resistant (pfmdr 1) gene that may be associated with anti-malarial drug resistance.


RESULTS
This assay has a sensitivity and specificity of 94% and 100% when compared to traditional PCR methods for genotyping. Only 54 of 68 (79%) paired pre- and post-culture DNA samples were concordant at all four loci.


CONCLUSIONS
Real-time PCR is a sensitive and specific method to detect SNP's in pfmdr 1. Genotypes of parasites after in vitro culture may not reflect that seen in vivo.
Purfield
Anne
A
Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill, NC, USA. purfield@med.unc.edu
Nelson
Amy
A
Laoboonchai
Anita
A
Congpuong
Kanungnij
K
McDaniel
Phillip
P
Miller
R Scott
RS
Welch
Kathy
K
Wongsrichanalai
Chansuda
C
Meshnick
Steven R
SR
D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D000818:Animals; D000962:Antimalarials / Q000494:pharmacology* / Q000627:therapeutic use; D016054:DNA, Protozoan / Q000032:analysis / Q000145:classification / Q000235:genetics; D018432:Drug Resistance, Multiple / Q000235:genetics; D019450:Genes, MDR / Q000235:genetics*; D005838:Genotype; D006801:Humans; D016778:Malaria, Falciparum / Q000188:drug therapy / Q000469:parasitology*; D010963:Plasmodium falciparum / Q000145:classification / Q000187:drug effects / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D015800:Protozoan Proteins / Q000235:genetics; D012680:Sensitivity and Specificity
D018528:ATP-Binding Cassette Transporters; D000962:Antimalarials; D016054:DNA, Protozoan; D015800:Protozoan Proteins; C059987:mdr gene protein, Plasmodium