Malaria Journal (May 2004)

A new method for detection of <it>pfmdr1 </it>mutations in <it>Plasmodium falciparum </it>DNA using real-time PCR

  • Welch Kathy,
  • Miller R Scott,
  • McDaniel Phillip,
  • Congpuong Kanungnij,
  • Laoboonchai Anita,
  • Nelson Amy,
  • Purfield Anne,
  • Wongsrichanalai Chansuda,
  • Meshnick Steven R

DOI
https://doi.org/10.1186/1475-2875-3-9
Journal volume & issue
Vol. 3, no. 1
p. 9

Abstract

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Abstract 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. Conclusion 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.