Immunology/Molecular Biology Core

免疫学/分子生物学核心

• 批准号: 7928162
• 负责人: MANUELA VERASTEGUI
• 金额: $ 16.25万
• 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/7928162
• 关键词: Animals; Antibodies; Antigens; Biological; Biological Assay; Biological Markers; Blood; Bolivia; Canis familiaris; Cavia; Centers for Disease Control and Prevention (U.S.); Chagas Disease; 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; innovation; 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将集中管理所有的免疫学和分子生物学 支持组成秘鲁TMRC的两个科学项目所需的程序，从而优化 资源的使用和保持严格的质量控制标准。核心的具体目标 B负责（1）协调与免疫学和分子生物学程序相关的所有工作， 研究项目和秘鲁TMRC的流行病学/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、执行和监测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 （5）为所获得的技术的有效转让和传播提供手段; (6)为大学生提供免疫学和寄生虫学培训的环境， 访问调查员。主要程序包括血清学和寄生虫学试验， 诊断人、豚鼠和狗的克氏锥虫感染（外鞭毛体抗原ELISA和 免疫荧光抗体试验、异种诊断、血培养）、使用锥虫 标准蛋白质印迹、ELISA、Falcon试验筛选试验中的排泄分泌抗原（TESA）- ELISA和一种新的现场友好的快速TESA-blot使用预先制备的条带，以允许快速转向- 克氏锥虫的分子生物学检测技术（检测PCR靶向核DNA卫星 重复序列或动基体DNA微环，核剪接前导序列阵列用于T.克鲁齐离散 分型单位，并区分它与T.ranglei，T.cruzi DNA RAPDS），微卫星位点 致病锥虫与表面增强激光解吸电离飞行时间质量 用于鉴定生物标志物的光谱法。所有即时免疫检测，快速- ELISA和TESA印迹的快速现场友好格式将在我们的现场进行， 阿雷基帕和圣克鲁斯，玻利维亚。PCR和RT-PCR也将在UPCH进行 该技术将被转移到玻利维亚。应变等新技术 多位点分型技术将在加州大学洛杉矶分校开发，然后转移到UPCH 实验室微卫星位点检测T. infestans将在CDC部门进行， 寄生虫病昆虫学实验室。人类、动物和T. 致病菌粪便和豚鼠的异种诊断，均将在阿雷基帕或 圣克鲁斯。所有的蛋白质组学工作将在英国伦敦帝国理工学院进行。的 这些程序的实施将支持这两个研究项目的科学工作 并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。 性能