Cloning of a fresh isolate of Plasmodium falciparum and drug sensitivity of the clones

  • Mirovsky P
  • Gay F
  • Bustos D
 et al. 
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Abstract

A freshly isolated strain of Plasmodium falciparum was cloned by limited dilution using a co-culture of infected erythrocytes on monolayers of functionally active rodent hepatocytes. 15 clones were isolated, and the anti-malarial activity of chloroquine, quinine, mefloquine and halofantrine against the clones, the original isolate, and a culture-adapted isolate was determined using a 48 h radioisotope microdilution method. The multiplication rates of all clones and the culture-adapted isolate were estimated by counting the number of parasitized cells on Giemsa-stained thin smears. Variations found in drug sensitivity and multiplication rate of different clones provided strong evidence of heterogeneity of a single strain parasite population. No morphological variation was detected by light microscopy. The use of a hepatocyte feeder layer improved the adaptation of cloned parasites to continuous culture conditions and thus enabled us to clone directly a fresh isolate, without losing clones during the culture-adaptation process. © 1990, Royal Society of Tropical Medicine and Hygiene. All rights reserved.

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Authors

  • Pavel Mirovsky

  • Frederick Gay

  • Dorina Bustos

  • Dominique Mazier

  • Marc Gentilini

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