1921
Volume 70, Issue 5
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

The influence of several factors on parasite growth and 50% inhibitory concentration (IC) for chloroquine was assessed. Most isolates stored at 4°C up to 72 hours grew when they were subsequently cultivated. However, parasite viability sharply decreased from 24 hours, and the mean chloroquine IC decreased significantly ( < 0.05). There was no evidence for selection of pre-culture populations due to storage alone. The time point when H-hypoxanthine was added (0 versus 18 hours) had no effect on the IC during the 42-hour incubation, but was associated with a lower IC when H-hypoxanthine was added after the initial 42-hour incubation during the 72-hour incubation. An increase in 3H-hypoxanthine incorporation and chloroquine IC was observed as the hematocrit was increased from 1.0% to 2.5%. For the same isolates, chloroquine IC values were generally similar when the initial parasitemia was between 0.1% and 0.5% but increased at higher (>0.75%) parasitemias. Based on these results, we recommend immediate cultivation after blood collection, a 42-hour incubation period with the addition of H-hypoxanthine at the beginning of incubation, a 1.5% hematocrit, and an initial parasitemia 0.1-0.5%. Further studies on serum substitutes, gas mixture, and comparison of isotopic and non-isotopic assays are needed to establish a standardized assay protocol.

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References

  1. Haynes JD, Diggs CL, Hines FA, Desjardins RE, 1976. Culture of human malaria parasites (P. falciparum). Nature 263 : 767–769. [Google Scholar]
  2. Trager W, Jensen JB, 1976. Human malaria parasites in continuous culture. Science 193 : 673–675. [Google Scholar]
  3. Rieckmann KH, Campbell GH, Sax LJ, Mrema JE, 1978. Drug sensitivity of Plasmodium falciparum: An in vitro microtechnique. Lancet 1 : 22–23. [Google Scholar]
  4. Wernsdorfer WH, 1980. Field evaluation of drug resistance in malaria. In vitro microtest. Acta Trop 37 : 222–227. [Google Scholar]
  5. Desjardins RE, Canfield CJ, Haynes JD, Chulay JD, 1979. Quantitative assessment of antimalarial activity in vitro by a semiautomated microdilution technique. Antimicrob Agents Chemother 16 : 710–718. [Google Scholar]
  6. Childs GE, Wimonwattrawatee T, Pooyindee N, 1988. Evaluation of an in vitro assay system for drug susceptibility of field isolates of Plasmodium falciparum from southern Thailand. Am J Trop Med Hyg 38 : 19–23. [Google Scholar]
  7. Pradines B, Mamfoumbi MM, Parzy D, Medang MO, Lebeau C, Mbina JRM, Doury JC, Kombila M, 1999. In vitro susceptibility of African isolates of Plasmodium falciparum from Gabon to pyronaridine. Am J Trop Med Hyg 60 : 105–108. [Google Scholar]
  8. Basco LK, 2002. Molecular epidemiology of malaria in Cameroon. XIII. Analysis of pfcrt mutations and in vitro chloroquine resistance. Am J Trop Med Hyg 67 : 388–391. [Google Scholar]
  9. Wernsdorfer WH, Payne D, 1988. Drug sensitivity tests in malaria parasites. Wernsdorfer WH, McGregor IA, eds. Malaria: Principles and Practice of Malariology. Volume 2. London: Churchill Livingstone Ltd., 1765–1800.
  10. Makler MT, Ries JM, Williams JA, Bancroft JE, Piper RC, Gibbins BL, Hinrichs DJ, 1993. Parasite lactate dehydrogenase as an assay for Plasmodium falciparum drug sensitivity. Am J Trop Med Hyg 48 : 739–741. [Google Scholar]
  11. Druilhe P, Moreno A, Blanc C, Brasseur PH, Jacquier P, 2001. A colorimetric in vitro drug sensitivity assay for Plasmodium falciparum based on a highly sensitive double-site lactate dehydrogenase antigen-capture enzyme-linked immunosorbent assay. Am J Trop Med Hyg 64 : 233–241. [Google Scholar]
  12. Noedl H, Wernsdorfer WH, Miller RS, Wongsrichanalai C, 2002. Histidine-rich protein II: a novel approach to malaria drug sensitivity testing. Antimicrob Agents Chemother 46 : 1658–1664. [Google Scholar]
  13. Doi H, Ishii A, Shimono K, 1988. A rapid in vitro assay system using anti-bromodeoxyuridine for drug susceptibility of Plasmodium falciparum. Trans R Soc Trop Med Hyg 82 : 190–193. [Google Scholar]
  14. Smeijsters LJJW, Zijlstra NM, Franssen FFJ, Overdulve JP, 1996. Simple, fast, and accurate fluorometric method to determine drug susceptibility of Plasmodium falciparum in 24-well suspension cultures. Antimicrob Agents Chemother 40 : 835–838. [Google Scholar]
  15. Mount DL, Nahlen BL, Patchen LC, Churchill FC, 1989. Adaptations of the Saker-Solomons test: Simple, reliable colorimetric field assays for chloroquine and its metabolites in urine. Bull World Health Organ 67 : 295–300. [Google Scholar]
  16. Jensen JB, 1988. In vitro cultivation of malaria parasites: erythrocytic stages. Wernsdorfer WH, McGregor IA, eds. Malaria: Principles and Practice of Malariology. Volume 1. London: Churchill Livingstone Ltd., 307–320.
  17. Basco LK, 2003. Molecular epidemiology of malaria in Cameroon. XV. Experimental studies on serum substitutes and supplements and alternative culture media for in vitro drug sensitivity assays using fresh isolates. Am J Trop Med Hyg 69 : 168–173. [Google Scholar]
  18. Basco LK, Tahar R, Escalante A, 2004. Molecular epidemiology of malaria in Cameroon. XVIII. Polymorphisms of the Plasmodium falciparum merozoite surface antigen-2 gene in isolates from symptomatic patients. Am J Trop Med Hyg 70: (in press).
  19. Ringwald P, Bickii J, Basco LK, 1996. In vitro activity of antima-larials against clinical isolates of Plasmodium falciparum in Yaoundé, Cameroon. Am J Trop Med Hyg 55 : 254–258. [Google Scholar]
  20. Basco LK, Ndounga M, Keundjian A, Ringwald P, 2002. Molecular epidemiology of malaria in Cameroon. IX. Characteristics of recrudescent and persistent Plasmodium falciparum infections after chloroquine or amodiaquine treatment in children. Am J Trop Med Hyg 66 : 117–123. [Google Scholar]
  21. Chulay JD, Haynes JD, Diggs CL, 1983. Plasmodium falciparum: assessment of in vitro growth by [3H]hypoxanthine incorporation. Exp Parasitol 55 : 138–146. [Google Scholar]
  22. Geary TG, Divo AA, Jensen JB, 1983. An in vitro assay system for the identification of potential antimalarial drugs. J Parasitol 69 : 577–583. [Google Scholar]
  23. Nivet C, Guillotte M, Pereira da Silva L, 1983. Plasmodium falciparum: one-step growth in a semi-defined medium and the stimulatory effect of human seric lipoproteins and liposomes. Exp Parasitol 55 : 147–151. [Google Scholar]
  24. Gluzman IY, Schlesinger PH, Krogstad DJ, 1987. Inoculum effect with chloroquine and Plasmodium falciparum. Antimicrob Agents Chemother 31 : 32–36. [Google Scholar]
  25. Duraisingh MT, Jones P, Sambou I, von Seidlein L, Pinder M, Warhurst DC, 1999. Inoculum effect leads to overestimation of in vitro resistance for artemisinin derivatives and standard an-timalarials: a Gambian field study. Parasitology 119 : 435–440. [Google Scholar]
  26. Basco LK, Ringwald P, 2001. Analysis of the key pfcrt point mutation and in vitro and in vivo response to chloroquine in Yaounde, Cameroon. J Infect Dis 183 : 1828–1831. [Google Scholar]
  27. Basco LK, 2003. Molecular epidemiology of malaria in Cameroon. XVI. Longitudinal surveillance of in vitro pyrimeth-amine resistance. Am J Trop Med Hyg 69 : 174–178. [Google Scholar]
  28. Langreth SG, Reese RT, Motyl MR, Trager W, 1979. Plasmodium falciparum: loss of knobs on the infected erythrocyte surface after long-term cultivation. Exp Parasitol 48 : 213–219. [Google Scholar]
  29. do Rosario VE, 1981. Cloning of naturally occurring mixed infections of malaria parasites. Science 212 : 1037–1038. [Google Scholar]
  30. Le Bras J, Deloron P, Ricour A, Andrieu B, Savel J, Coulaud JP, 1983. Plasmodium falciparum: drug sensitivity in vitro of isolates before and after adaptation to continuous culture. Exp Parasitol 56 : 9–14. [Google Scholar]
  31. Ringwald P, Meche FS, Bickii J, Basco LK, 1999. In vitro culture and drug sensitivity assay of Plasmodium falciparum with non-serum substitute and acute-phase sera. J Clin Microbiol 37 : 700–705. [Google Scholar]
  32. Basco LK, 2003. Molecular epidemiology of malaria in Cameroon. XVII. Baseline monitoring of atovaquone-resistant Plasmodium falciparum by in vitro drug assays and cytochrome b gene sequence analysis. Am J Trop Med Hyg 69 : 179–183. [Google Scholar]
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  • Received : 13 Jul 2003
  • Accepted : 08 Jan 2004

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