Creating and Validating Ligase Primers to Detect Single Nucleotide Polymorphisms Associated with Atovaquone Resistance in Plasmodium falciparum

Muharib Alruwaili, Rubayet Elahi, Donelly van Schalkwyk, Colin Sutherland, Theresa Shapiro, Sean Prigge, David Sullivan

Research output: Contribution to journalArticlepeer-review

Abstract

Atovaquone-proguanil is one of the most commonly prescribed malaria prophylactic drugs. However, sporadic mutations conferring resistance to atovaquone have been detected in recent years associated with single nucleotide polymorphisms (SNPs) in the Plasmodium falciparum cytochrome b ( pfcytb) gene. Monitoring polymorphisms linked with resistance is essential in assessing the prevalence of drug resistance and may help in designing strategies for malaria control. Several approaches have been used to study genetic polymorphisms associated with antimalarial drug resistance. However, they either lack high throughput capacity or are expensive in time or money. Ligase detection reaction fluorescent microsphere assay (LDR-FMA) provides a high-throughput method to detect genetic polymorphisms in P. falciparum. In this study, we have created primers to detect SNPs associated with clinically relevant atovaquone resistance using LDR-FMA and validated them in clinical samples. Four SNPs from pfcytb gene were analyzed using LDR-FMA. The results were 100% consistent with DNA sequence data, indicating that this method has potential as a tool to detect genetic polymorphisms associated with atovaquone resistance in P. falciparum.

Original languageEnglish (US)
Pages (from-to)777-782
Number of pages6
JournalThe American journal of tropical medicine and hygiene
Volume108
Issue number4
DOIs
StatePublished - Apr 5 2023

ASJC Scopus subject areas

  • Infectious Diseases
  • Virology
  • Parasitology

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