Enhanced ex vivo Plasmodium vivax intraerythrocytic enrichment and maturation for rapid and sensitive parasite growth assays

Gabriel W. Rangel, Martha A. Clark, Usheer Kanjee, Caeul Lim, Kathryn Shaw-Saliba, Maria José Menezes, Anjali Mascarenhas, Laura Chery, Edwin Gomes, Pradipsinh K. Rathod, Marcelo U. Ferreira, Manoj T. Duraisingh

Research output: Contribution to journalArticlepeer-review

25 Scopus citations


Plasmodium vivax chloroquine resistance has been documented in nearly every region where this malaria-causing parasite is endemic. Unfortunately, P. vivax resistance surveillance and drug discovery are challenging due to the low parasitemias of patient isolates and poor parasite survival through ex vivo maturation that reduce the sensitivity and scalability of current P. vivax antimalarial assays. Using cryopreserved patient isolates from Brazil and fresh patient isolates from India, we established a robust enrichment method for P. vivax parasites. We next performed a medium screen for formulations that enhance ex vivo survival. Finally, we optimized an isotopic metabolic labeling assay for measuring P. vivax maturation and its sensitivity to antimalarials. A KCl Percoll density gradient enrichment method increased parasitemias from smallvolume ex vivo isolates by an average of > 40-fold. The use of Iscove's modified Dulbecco's medium for P. vivax ex vivo culture approximately doubled the parasite survival through maturation. Coupling these with [3H]hypoxanthine metabolic labeling permitted sensitive and robust measurements of parasite maturation, which was used to measure the sensitivities of Brazilian P. vivax isolates to chloroquine and several novel antimalarials. These techniques can be applied to rapidly and robustly assess the P. vivax isolate sensitivities to antimalarials for resistance surveillance and drug discovery.

Original languageEnglish (US)
Article numbere02519-17
JournalAntimicrobial agents and chemotherapy
Issue number4
StatePublished - Apr 2018


  • Antimalarial agents
  • Antimalarials
  • Chloroquine
  • Malaria
  • Plasmodium vivax

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)
  • Infectious Diseases


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