Evaluation of Plasmodium vivax Genotyping Markers for Molecular Monitoring in Clinical Trials
Evaluation of Plasmodium vivax Genotyping Markers for Molecular Monitoring in Clinical Trials
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DOI：
10.1086/597303
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发表时间：
2009-04-01
影响因子：
6.4
通讯作者：
Felger, Ingrid
中科院分区：
文献类型：
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作者：
Koepfli, Cristian;Mueller, Ivo;Felger, Ingrid
背景许多抗疟干预措施都伴随着对寄生虫感染的分子监测，并使用了基于不同多态性标记基因的一些分子分型技术。在这里，我们描述了一种基因分型技术，提供了一种快速，精确的方法来研究间日疟原虫感染动态的情况下，必须遵循随着时间的推移，个别克隆。用来自体内药物功效研究的样品测试该方法。采用毛细管电泳技术对来自巴布亚新几内亚3个村庄的93-108份间日疟原虫阳性血样中9个潜在遗传标记（5个裂殖子表面蛋白[msp]基因和4个微卫星）的PCR扩增片段大小进行了分析。微卫星MS 16和Pv3.27在研究区表现出最大的多样性，分别有66和31个不同的等位基因，其次是2个片段的msp 1和2个其他微卫星。msp 3 α、msp 4和msp 5显示有限的多态性。即使对于最多样化的标记，最高等位基因频率也达到6%（MS 16）或13%（Pv3.27）。为了降低与基线检测到的单倍型相同的寄生虫重复感染的理论概率，我们建议联合收割机组合至少2种标记物用于对个体间日疟原虫感染进行基因分型。
Background. Many antimalarial interventions are accompanied by molecular monitoring of parasite infections, and a number of molecular typing techniques based on different polymorphic marker genes are used. Here, we describe a genotyping technique that provides a fast and precise approach to study Plasmodium vivax infection dynamics during circumstances in which individual clones must be followed over time. The method was tested with samples from an in vivo drug efficacy study.Methods. The sizes of polymerase chain reaction fragments were evaluated by capillary electrophoresis to determine the extent of size polymorphism for 9 potential genetic markers (5 genes of merozoite surface proteins [msp] and 4 microsatellites) in 93-108 P. vivax-positive blood samples from 3 villages in Papua New Guinea.Results. The microsatellites MS16 and Pv3.27 showed the greatest diversity in the study area, with 66 and 31 different alleles, respectively, followed by 2 fragments of msp1 and 2 other microsatellites. msp3 alpha, msp4, and msp5 revealed limited polymorphism.Conclusions. Even for the most diverse markers, the highest allelic frequencies reached 6% (MS16) or 13% (Pv3.27). To reduce the theoretical probability of superinfection with parasites that have the same haplotype as that detected at baseline, we propose to combine at least 2 markers for genotyping individual P. vivax infections.