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
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
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DOI：
10.4269/ajtmh.2006.75.777
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发表时间：
2006-11-01
影响因子：
3.3
通讯作者：
Basco, Leonardo K.
中科院分区：
文献类型：
--
作者：
Basco, Leonardo K.
随着许多疟疾控制计划放弃单一疗法而求助于联合疗法，恶性疟原虫野外分离株的体外药物反应和耐药性分子标记的相关性研究变得越来越重要。体外药敏试验的标准化和优化是分子标记物在该领域验证的先决条件之一。本研究旨在评估和比较新鲜获得的分离株在不同培养基中至少第一个红细胞周期的生长情况，并确定在替代培养基中对氯喹的体外反应。寄生虫在Dulbecco改良Eagle培养基（DME）-人血清、Iscove改良Dulbecco培养基（IMDM）-人血清、RPMI 1640培养基-山羊血清和含有IMDM和F-12的1：1（v/v）混合物（补充成年牛血清的硫酸铵组分）的无血清培养基中的生长始终高于RPMI 1640培养基-人血清混合物。在补充人血清的DME、IMDM和RPMI 1640培养基中测定的氯喹反应水平与对照（RPMI 1640-人血清）无显著差异（P > 0.05）。这项研究表明，替代媒体可用于优化寄生虫的生长在关键的初始阶段的过渡，从体内到体外条件。这些媒体的能力，以支持长期培养的恶性疟原虫需要进一步调查。
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.