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A new method for detection of pfmdr1mutations in Plasmodium falciparumDNA using real-time PCR

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

File Type: pdf | Filesize: 277.1 KB | Date Added: 2012-09-05 | Date Created: 2004-05-07

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.