Development of a Field Applicable Screening Tool (FAST) kit for detecting resista

开发用于检测耐药性的现场适用筛选工具 (FAST) 套件

• 批准号: 7657009
• 负责人: Martin James Donnelly
• 金额: $ 43.63万
• 依托单位: TMLIVERPOOL SCHOOL OF TROPICAL MEDICINE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7657009
• 关键词: Address; Affect; Africa South of the Sahara; African; Alleles; Anopheles Genus; Anopheles gambiae; Biological Assay; Candidate Disease Gene; Carbamates; Collaborations; Country; Culicidae; Custom; DNA; Development; Diagnostic; Disease; Failure; Filariasis; Frequencies; Generic Drugs; Genes; Genetic Markers; Genetic Transcription; Genome; Goals; Insecticide Resistance; Insecticides; Laboratories; Malaria; Maps; Meta-Analysis; Metabolic; Molecular; Monitor; Mutation; Phase; Population; Program Development; Public Health; Regulator Genes; Research; Research Personnel; Residual state; Resistance; Sampling; Screening procedure; Site; Testing; Tissue-Specific Gene Expression; Transcript; Tropical Disease; Validation; Variant; World Health Organization; base; cyhalothrin; design; dichlorodiphenyltrichloroethane; filaria; innovation; programs; pyrethroid; resistance mechanism; success; tool; vector control

DESCRIPTION (provided by applicant): The long term objective is to facilitate the rational use of insecticides in malaria and filaria control campaigns based by upon a more complete understanding of resistance mechanisms. The specific aims of this proposal are to: i) identify genes that render Anopheles gambiae s.l. mosquitoes resistant to a range of insecticides that are used for indoor residual spraying (IRS) for malaria and filariasis namely an organochloride (DDT), a carbamate (Bendiocarb) and a class II pyrethroid (?-cyhalothrin). ii) develop DNA-based insecticide resistance screening tools for use by the staff of vector control programmes in the malaria and filariasis affected countries of sub-Saharan Africa. These routine resistance screens will permit routine monitoring of mutations giving information vital for predicting the success of IRS and ITN programmes. The objective will be addressed in three specific phases: Phase 1 - The discovery of resistance-associated loci using candidate gene association mapping and whole genome microarrays. In two focal African countries, genes potentially involved in resistance to three major IRS insecticides (DDT, Bendiocarb, ?-cyhalothrin) will be identified using two complementary approaches:- i) Variation at the DNA level will be investigated using a custom candidate gene 1536plex SNP array and ii) differential gene expression between resistant and susceptible Anopheles gambiae will be investigated via whole genome microarrays. Putative resistance-associated SNPs within and around the most promising genes or gene regulators will be taken forward to phase 2 for wider field validation. Phase 2 - Candidate gene validation in the PMI and TDR network countries of sub-Saharan Africa. The putative resistance-associated SNPs identified in phase 1 will be screened for association with resistance in a much larger sample from a large section of sub-Saharan Africa via collaboration with extant World Health Organization/Tropical Disease Research (TDR) and President's Malaria Initiative (PMI) programmes. The aim is to screen for association of SNPs with resistance within and across populations producing a multi-site association meta-analysis to identify resistance-associated SNPs of major and/or generic effect. Phase 3 - Integrating metabolic resistance markers into a Field Applicable Screening Tool. The final phase of the programme will involve design, testing and roll-out of a field applicable screening tool (FAST), which will comprise straightforward and robust diagnostics to be used in laboratories in the disease-affected countries. The FAST diagnostics will include the most important genetic markers for resistance to the IRS insecticides, in addition to any critical pre-identified markers (e.g. for species/ molecular form identification). FAST roll-out will occur via collaboration with extant Innovative Vector Control Consortium (IVCC), TDR and PMI programmes. The development of insecticide resistance in Anopheles mosquitoes is a major threat to malaria control. Conventional means of assessing resistance are costly, insensitive and inaccurate. We propose to develop assays that will allow disease programme managers to detect resistance when it is still at a low frequency and before control failure has occurred.

描述（由申请人提供）：长期目标是在更全面地了解耐药机制的基础上，促进疟疾和丝虫控制活动中杀虫剂的合理使用。该提案的具体目标是： i) 鉴定使冈比亚按蚊 s.l. 产生的基因。蚊子对用于治疗疟疾和丝虫病的室内滞留喷洒 (IRS) 的一系列杀虫剂具有抗药性，这些杀虫剂包括有机氯化物 (DDT)、氨基甲酸酯 (Bendiocarb) 和 II 类拟除虫菊酯 (β-三氯氟氰菊酯)。 ii) 开发基于 DNA 的杀虫剂耐药性筛查工具，供受疟疾和丝虫病影响的撒哈拉以南非洲国家病媒控制项目的工作人员使用。这些常规耐药性筛查将允许对突变进行常规监测，为预测 IRS 和 ITN 计划的成功提供至关重要的信息。 该目标将分三个具体阶段来实现： 第一阶段 - 使用候选基因关联作图和全基因组微阵列发现耐药性相关位点。在两个非洲重点国家，将使用两种互补方法来鉴定可能与三种主要 IRS 杀虫剂（DDT、苯灭威、β-三氟氯氰菊酯）抗性相关的基因：- i) 将使用定制候选基因 1536plex 来研究 DNA 水平的变异SNP阵列和ii)抗性和易感冈比亚按蚊之间的差异基因表达将通过全基因组微阵列进行研究。最有希望的基因或基因调节因子内部和周围的假定的抗性相关 SNP 将进入第二阶段，以进行更广泛的领域验证。 第 2 阶段 - 在撒哈拉以南非洲的 PMI 和 TDR 网络国家进行候选基因验证。通过与现有的世界卫生组织/热带病研究 (TDR) 和总统疟疾倡议的合作，将在第一阶段中确定的假定的与耐药性相关的 SNP 在来自撒哈拉以南非洲大部分地区的更大样本中筛选与耐药性的关联。 PMI）计划。目的是筛选人群内部和人群之间 SNP 与耐药性的关联，进行多位点关联荟萃分析，以确定主要和/或一般效应的耐药性相关 SNP。 第 3 阶段 - 将代谢抗性标记整合到现场适用的筛选工具中。该计划的最后阶段将涉及设计、测试和推出现场适用的筛查工具（FAST），该工具将包括在受疾病影响国家的实验室中使用的简单而强大的诊断方法。除了任何关键的预先识别标记（例如用于物种/分子形式识别）之外，FAST 诊断还将包括最重要的 IRS 杀虫剂抗性遗传标记。 FAST 的推出将通过与现有创新病媒控制联盟 (IVCC)、TDR 和 PMI 计划的合作进行。 按蚊对杀虫剂耐药性的发展是疟疾控制的主要威胁。评估耐药性的传统方法成本高昂、不敏感且不准确。我们建议开发一些检测方法，使疾病项目管理者能够在耐药性仍处于低频率时且在控制失败发生之前检测到耐药性。