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MOLECULAR EPIDEMIOLOGY OF MALARIA IN CAMEROON. XXIII. EXPERIMENTAL STUDIES ON SERUM SUBSTITUTES AND ALTERNATIVE CULTURE MEDIA FOR IN VITRO DRUG SENSITIVITY ASSAYS USING CLINICAL ISOLATES OF PLASMODIUM FALCIPARUM

Correlation studies on the in vitro drug response of field isolates of Plasmodium falciparum and molecular markers for drug resistance are becoming important as many malaria control programs abandon monotherapies and resort to combination therapies. The standardization and optimization of the in vitro drug sensitivity assay are one of the prerequisites for validating molecular markers in the field. The present study was designed to assess and compare the growth of freshly obtained isolates for at least the first erythrocytic cycle in various culture media and determine the in vitro response to chloroquine in alternative media. Parasite growth was consistently higher in Dulbecco’s modified Eagle’s medium (DME)–human serum, Iscove’s modified Dulbecco’s medium (IMDM)–human serum, RPMI 1640 medium–goat serum, and a serum-free medium containing 1:1 (v/v) mixture of IMDM and F-12 supplemented with an ammonium sulfate fraction of adult bovine serum than in RPMI 1640 medium–human serum mixture. The level of chloroquine response determined in human serum-supplemented DME, IMDM, and RPMI 1640 media did not differ significantly (P > 0.05) from the control (RPMI 1640–human serum). This study suggests that alternative media may be used to optimize parasite growth during the critical initial phase of transition from in vivo to in vitro conditions. The capacity of these media to support long-term cultivation of P. falciparum requires further investigation.

Received May 1, 2005. Accepted for publication August 2, 2006.

Acknowledgment: I thank the personnel of the Nlongkak Catholic missionary dispensary for their aid in recruiting patients.

Financial support: This study was supported by the French Ministry of Research (Programme PAL+).

^* Address correspondence to Leonardo K. Basco, Laboratoire de Recherche sur le Paludisme, Organisation de Coordination pour la lutte contre les Endémies en Afrique Centrale, BP 288, Yaounde, Cameroon. E-mail: lkbasco{at}yahoo.fr

Author’s address: Leonardo K. Basco, Unité de Recherche Paludologie Afro-Tropicale, Institut de Recherche pour le Développement and Laboratoire de Recherche sur le Paludisme, Organisation de Coordination pour la lutte contre les Endémies en Afrique Centrale, BP 288, Yaounde, Cameroon.

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