Immunology/Molecular Biology Core

免疫学/分子生物学核心

• 批准号: 8129636
• 负责人: MANUELA VERASTEGUI
• 金额: $ 16.27万
• 依托单位: UNIVERSIDAD PERUANA CAYETANO HEREDIA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8129636
• 关键词: Animals; Antibodies; Antigens; Biological; Biological Assay; Biological Markers; Blood; Bolivia; Canis familiaris; Cavia; Centers for Disease Control and Prevention (U.S.); Collection; DNA; Detection; Diagnosis; Diffusion; Entomology; Environment; Enzyme-Linked Immunosorbent Assay; Epidemiology; Equipment and Supplies; Feces; Human; Immunologic Tests; Immunology; Infection; Laboratories; Laboratory Personnel; London; Mass Spectrum Analysis; Microsatellite Repeats; Molecular; Molecular Biology; Monitor; Nuclear; Parasitic Diseases; Parasitology; Performance; Peru; Procedures; Proteomics; Research Personnel; Research Project Grants; Resources; Satellite DNA; Scientist; Screening procedure; Serological; Site; Specimen; Spliced Leader Sequences; Techniques; Technology; Testing; Time; Training; Triatoma; Trypanosoma cruzi; Visit; Western Blotting; Work; Xenodiagnosis; college; data management; kDNA Minicircles; novel; point of care; surface enhanced laser desorption ionization; university student

Core B will centralize the management of all of the immunology and molecular biology procedures neededto support the two scientific projects that form the Peru TMRC, thus optimizing the use of resources and maintaining rigorousquality control standards. The specific aims of Core B are to (1) coordinate all work related to immunology and molecular biology procedures between both Research Projects and the Epidemiology/GIS/Data Management core of the Peru TMRC; (2) coordinate and oversee collection and handling procedures for all biological specimens; (3) standardize, perform, and monitor all immunological and molecular biology procedures; (4) optimize the use of laboratory personnel, equipment and supplies; (5) coordinate the register and collation of assay results; (5) provide the means for an efficient transfer and diffusion of acquired technology; (6) provide the environment for immunology and parasitology training for University students and visiting investigators. The major procedures consist of serologic and parasitologic assays for the diagnosis of T.cruzi infection in humans, guinea pigs and dogs (epimastigote antigen ELISA and immunofluoresecent antibody tests, xenodiagnosis, hemoculture), the use of Trypomastigote excreted-secreted antigens (TESA) in standard Western blot, ELISA, Falcon assay screening test - ELISA and a novel field-friendly rapid TESA-blot using pre-prepared strips to allow rapid turn- around, a range of T.cruzi molecular techniques (Detection PCR targeting nuclear DNA satellite repeats or kinetoplast DNA minicircles, Nuclear spliced leader array for classifying T. cruzi discrete typing units' and distinguishing it from T.ranglei, T.cruzi DNA RAPDS), microsatellite loci for Triatoma infestans and Surface-enhanced laser desorption ionization-time of flight mass spectrometry for the indentification of biomarkers. All point-of-care immunological tests, FAST- ELISA and the rapid field-friendly format of the TESA blot will be conducted in our field sites, Arequipa and Santa Cruz, Bolivia. PCRand RT- PCR will also be performed at the UPCH laboratory and then the technique will be transferred to Bolivia. New techniques such as strain typing using multi - locus typing will be developed at UCLA andthen transferred to the UPCH laboratory. Microsatellite loci testing for T. infestans will be performed at the CDC Division of Parasitic Diseases entomology laboratory. Hemoculture of blood from humans, animals and T. infestans feces, and xenodiagnosis in guinea pigs, will all be performed on site either in Arequipa or Santa Cruz. All proteomics work will be conducted at the Imperial College, London, UK. The implementation of these procedures will support the scientific work of the two Research Projects and provide significant training opportunities for Peruvian scientists through interchanges with the US, UK and Brazilian collaborators and consultants. PERFORMANCE.

核心 B 将集中管理所有免疫学和分子生物学 支持构成秘鲁 TMRC 的两个科学项目所需的程序，从而优化 使用资源并维持严格的质量控制标准。核心的具体目标 B 是 (1) 协调与免疫学和分子生物学程序相关的所有工作 秘鲁 TMRC 的研究项目和流行病学/GIS/数据管理核心； (2) 协调和监督所有生物样本的收集和处理程序； (3) 标准化、执行和监控所有免疫学和分子生物学程序； (4)优化 实验室人员、设备和用品的使用； (5) 协调登记和整理 化验结果； (5) 提供有效转让和传播所获得技术的手段； (6)为大学生提供免疫学和寄生虫学培训的环境 来访的调查人员。主要程序包括血清学和寄生虫学检测 人类、豚鼠和狗中克氏锥虫感染的诊断（表鞭毛体抗原 ELISA 和 免疫荧光抗体测试、异种诊断、血液培养）、锥鞭毛体的使用 标准蛋白质印迹、ELISA、Falcon 测定筛选试验中的分泌-分泌抗原 (TESA) - ELISA 和一种新颖的现场友好型快速 TESA 印迹，使用预先准备的试纸条实现快速翻转 周围，​​一系列 T.cruzi 分子技术（检测 PCR 靶向核 DNA 卫星 重复序列或动质体 DNA 小环，核剪接前导阵列，用于对 T. cruzi 离散进行分类 分型单位'并将其与 T.ranglei、T.cruzi DNA RAPDS 区分开来，微卫星位点 马铃薯锥蝽和表面增强激光解吸-飞行质量电离时间 光谱测定法用于鉴定生物标志物。所有即时免疫学测试，FAST- ELISA 和 TESA 印迹的快速现场友好格式将在我们的现场进行， 玻利维亚阿雷基帕和圣克鲁斯。 PCR 和 RT-PCR 也将在 UPCH 进行 实验室，然后该技术将转移到玻利维亚。应变等新技术 使用多基因座分型的分型将在加州大学洛杉矶分校开发，然后转移到 UPCH 实验室。 T. infestans 的微卫星位点检测将在 CDC 部门进行 寄生虫病昆虫学实验室。对人类、动物和 T. 的血液进行血液培养。 致病菌粪便和豚鼠异种诊断都将在阿雷基帕或阿雷基帕现场进行 圣克鲁斯。所有蛋白质组学工作将在英国伦敦帝国理工学院进行。这 这些程序的实施将支持两个研究项目的科学工作 通过与秘鲁科学家的交流，为秘鲁科学家提供重要的培训机会 美国、英国和巴西的合作者和顾问。 表现。