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

• Published in: Malaria Journal
• Main Authors: Welch Kathy, Miller R Scott, McDaniel Phillip, Congpuong Kanungnij, Laoboonchai Anita, Nelson Amy, Purfield Anne, Wongsrichanalai Chansuda, Meshnick Steven R
• Format: Article in Journal/Newspaper
• Language: English
• Published: BMC 2004
• Subjects: Arctic medicine. Tropical medicine; RC955-962; Infectious and parasitic diseases; RC109-216; Arctic
• Online Access: https://doi.org/10.1186/1475-2875-3-9; https://doaj.org/article/c3b4058a04534c7fb34599f5b25da1fd

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 .