Pathogenesis and Diagnosis

项目总结(见说明)： 恶性疟原虫表现出丰富的基因多样性，由宿主和媒介免疫、药物 压力，环境变化？是它作为一种持久病原体成功的关键因素。理解 基因多样性的性质、范围和分布以及它如何随时间变化将是设计 最有效和最有效的疟疾控制措施。到目前为止，还没有关于人口的数据 非洲南部地区恶性疟原虫的基因分布。研究区C的总体目标是 在ICEMR研究地点建立恶性疟原虫种群基因的区域分布。这将是 使用基于阵列和基于聚合酶链式反应的方法完成，这些方法将提供高分辨率和低分辨率 寄生虫遗传多样性概况。除了基于种群的问题外，已探明的裂殖子表面 蛋白-2(Msp2)基因分型试验将用于评估个体内寄生虫的多样性和特殊的 强调低传播率地区的无症状/无症状感染。目标1)获得高分辨率 ICEMR研究点内和点间寄生虫分离株的基因差异 确定寄生虫遗传多样性的性质和范围。一种高密度的恶性疟原虫瓷砖阵列 这些研究将使用Affymetrix平台。目的2)实现并提炼基于PCR的条形码 作为一种简单、经济有效的工具来粗略监测寄生虫遗传结构的变化 ICEMR研究地点的人口。条形码将由实时RT-PCR定义的-25个SNP组成。 虽然条形码将提供比基于阵列的方法更低的分辨率基因，但其更低的成本和 易用性将使人们能够更全面地了解地区人口结构，这将 便于识别由于控制措施或其他因素而导致的随时间变化。目标3)确定 低传播率和高传播率人群中寄生虫克隆多样性的水平和动态 ICEMR研究地点的区域。将根据检测到的多氟氯烃来评估感染性氟龙(MOI) 聚合酶链式反应检测msp2基因的多态性。将特别强调无提示和无提示 以及这些个体在传播过程中可能扮演的角色。令人期待的是 基因分型和MOI数据将被用来建立预测遗传多样性对 传播动力学、药物治疗努力和发病机制。

PROJECT SUMMARY (See instructions): The abundant geneflc diversity exhibited by P. falciparum, shaped by host and vector immunity, drug pressure, environmental change? is a key element to its success as a persistent pathogen. Understanding the nature, extent and distribution of geneflc diversity and how it changes over time will be key to devising the most efflcient and effective control measures for malaria. To date, there is no data on the population geneflcs of P. falciparium in the southern region of Africa. The overall goal of Research Area C is to establish regional proflles of the populaflon geneflcs of P. falciparium in the ICEMR study sites. This will be accomplished using array-based and PCR-based approaches that will provide both high- and low-resoluflon profiles of parasite genetic diversity. In addition to population-based issues, the proven merozoite surface protein-2 (msp2) genotyping assay will be used to assess intra-individual parasite diversity with special emphasis in asymptomatic/silent infection in areas of low transmission. Aim 1) Obtain a high-resolution profile ofthe genotypic differences between parasite isolates within and behween ICEMR study sites in order to establish the nature and scope of parasite genetic diversity. A high-density P. falciparum tiling array on the Affymetrix platform will be used for these studies. Aim 2) Implement and refine a PCR-based barcode approach as a simple, cost-effective tool for roufinely monitoring changes in the genetic structure of parasite populations in the ICEMR study sites. The barcode will consist of -25 SNPs defined by real time RT PCR. While the barcode will provide a lower resolution genotype than the array-based approach, its lower cost and ease of use will allow a much more comprehensive profile of regional populafion geneflc structure that will facilitate the identiflcation of changes over time due to control measures or other factors. Aim 3) Determine the level and dynamics of parasite clonal diversity within individuals residing in low- and high-transmission areas in the ICEMR study sites. Mulflplicity of infecflon (MOI) will be assessed based on the detecflon of polymorphisms in the msp2 gene by PCR. Special emphasis will be placed on asymptomaflc and silent infecflons and the role that these individuals may have in transmission. The expectaflon is that the genotyping and MOI data will be used to create models for predicflon of geneflc diversity influence upon transmission dynamics, drug treatment efforts, and pathogenesis.