Establishment and evaluation of a qPCR method for the detection of pfmdr1 mutations in Plasmodium falciparum, the causal agent of fatal malaria

Diagn Microbiol Infect Dis. 2024 Sep;110(1):116400. doi: 10.1016/j.diagmicrobio.2024.116400. Epub 2024 Jun 20.

Abstract

Drug resistance surveillance is a major integral part of malaria control programs. Molecular methods play a pivotal role in drug resistance detection and related molecular research. This study aimed to develop a rapid and accurate detection method for drug resistance of Plasmodium falciparum (P. falciparum). A quantitative real-time PCR (qPCR) assay has been developed that identifies the mutation at locus A256T in the P.falciparum multi-drug resistance(pfmdr1) gene producing amino acid change at position 86. The results of 198 samples detected by qPCR were consistent with nested PCR and sequencing, giving an accuracy of 94.3%. The sensitivity, specificity, positive and negative predictive value of qPCR were 85.7%, 97.6%, 90.0% and 96.4%, respectively. The results of qPCR are basically consistent with the nested PCR, which is expected to replace the nested PCR as a new molecular biological method for drug resistance detection, providing reliable technical support for global malaria prevention and control.

Keywords: Antimalarial drug resistance; Molecular assays; Nested PCR; Plasmodium falciparum; Quantitative real-time PCR (qPCR); Surveillance.

Publication types

  • Evaluation Study

MeSH terms

  • Antimalarials / pharmacology
  • Antimalarials / therapeutic use
  • Drug Resistance / genetics
  • Humans
  • Malaria, Falciparum* / diagnosis
  • Malaria, Falciparum* / parasitology
  • Multidrug Resistance-Associated Proteins* / genetics
  • Mutation
  • Plasmodium falciparum* / genetics
  • Real-Time Polymerase Chain Reaction* / methods
  • Sensitivity and Specificity*

Substances

  • Mdr1 protein, Plasmodium falciparum
  • Multidrug Resistance-Associated Proteins
  • Antimalarials