Pathogen genotyping in polyclonal infections: Application of a fluorogenic polymerase-chain-reaction assay in malaria
Pathogen genotyping in polyclonal infections: Application of a fluorogenic polymerase-chain-reaction assay in malaria
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DOI：
10.1086/378521
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发表时间：
2003-10-15
影响因子：
6.4
通讯作者：
Dujardin, JC
中科院分区：
文献类型：
--
作者：
Decuypere, S;Elinck, E;Dujardin, JC
多克隆感染的病原体基因分型受到两个主要缺点的限制：（1）如何确定1个基因中检测到的多个突变是否属于同一克隆；（2）如何评估给定样本中不同基因型的比例。对于恶性疟原虫疟疾的耐药基因分型，我们通过使用荧光测定来解决这些问题，该测定结合了探针上荧光团和聚合酶链反应引物之间的荧光共振能量转移以及熔解曲线分析。我们证明，该工具可以更准确地了解复杂生物样本中存在的恶性疟原虫种群。
Pathogen genotyping of polyclonal infections is limited by 2 major drawbacks: (1) how to establish whether multiple mutations detected in 1 gene belong to the same clone and (2) how to evaluate the proportion of different genotypes in a given sample. For drug-resistance genotyping in Plasmodium falciparum malaria, we address these problems by using a fluorogenic assay that combines fluorescence-resonance energy transfer, between fluorophores present on a probe and a polymerase-chain-reaction primer, and a melt-curve analysis. We demonstrate that this tool allows a more accurate insight into the P. falciparum populations present in complex biological samples.