Improving diagnostic capabilities for typhoid fever

描述(申请人提供)：目前，在世界伤寒流行地区，尚无良好的伤寒(或携带)诊断试验。这一缺陷使适当抗菌剂的靶向给药复杂化，并且是更广泛地支持伤寒控制和疫苗计划的主要障碍。在这个分两个阶段的R21(探索)/R33(扩展和发展)提案中，我们建立在持续的国际合作努力的基础上，以支持以下具体目标和里程碑：R21：第一年和第二年：创新假设驱动的项目1.抗S-S的评估。伤寒免疫球蛋白A ALS体液检测系统作为伤寒诊断。2.尿液酶联免疫吸附试验或试纸法的建立。3.筛选无症状载体血清，寻找针对伤寒沙门氏菌体内表达和载体状态所特有的抗体。在第二年年底(R21)，我们建议(1)开发至少一种原型方法(基于ALS或基于尿液)，用于孟加拉国达卡的后续评估，以及(2)筛选携带者血液，以评估是否存在独特的信号。R33：第3-5年：额外的创新探索性研究和诊断学的扩展开发4.在孟加拉国达卡(流行区)进行的R21阶段的现场(流行区)测试原型分析(肌萎缩侧索硬化症和/或尿器[S])，如果有希望的话。5.先进的运营商诊断，如果有希望的话。如果没有希望：6.进一步探索使用微流控甘露糖结合凝集素“捕获”系统来提高急性感染患者血液中伤寒沙门氏菌的回收率。7.探索伤寒的干扰素-γ检测方法。8.探索诊断检测伤寒患者血液中伤寒沙门氏菌m RNA/c DNA(相对于gDNA)的能力，以提高敏感性。到第5年年底的主要交付成果(R33)：开发比流行区目前的分析更敏感和更特异的诊断方法(我们的初始目标将是&gt；90%的灵敏度；&gt；90%的特异度；比目前在流行区可用的/正在使用的分析有显著的改进)。

DESCRIPTION (provided by applicant): Currently, there is no good diagnostic test for typhoid fever (or carriage) in typhoid endemic areas of the world. This deficiency complicates the targeted administration of appropriate antimicrobials, and is a major impediment to the more widespread endorsement of typhoid control and vaccine programs. In this two-stage R21 (exploration)/R33 (expansion and development) proposal, we build upon an ongoing international collaborative effort to support the following specific aims and milestones: R21: years 1 and 2: innovative hypothesis-driven projects 1. Evaluation of anti-S. typhi IgA ALS fluid detection system as a typhoid diagnostic. 2. Development of urine ELISA or dipstick assay. 3. Screen asymptomatic carrier serum for antibodies targeting S. typhi antigens expressed in vivo and unique to the carrier state. Main milestones by the end of year 2 (R21), we propose (1) to have developed at least one prototype approach (either ALS-based or urine-based) to use in subsequent evaluations in Dhaka, Bangladesh, and (2) to have screened carrier blood to assess if a unique signal is present. R33: years 3-5: additional innovative exploratory research and expanded development of diagnostics 4. Field (endemic zone) test prototype assays in Dhaka, Bangladesh (ALS and/or urine device[s]) from R21 phase, if promising. 5. Advance carrier diagnostic, if promising. If not promising: 6. Further explore the use of a microfluidic mannose binding lectin "capture" system to increase recovery of S. typhi in the blood of acutely infected patients. 7. Explore interferon-gamma based detection assay (IGA) approach for typhoid. 8. Explore the ability to diagnostically detect S. typhi mRNA/cDNA (as opposed to gDNA) in the blood of infected humans with typhoid, to improve sensitivity. Main deliverable by the end of year 5 (R33): development of diagnostic approach(es) that is/are more sensitive and specific than current assays in an endemic zone (our initial goal will be >90% sensitivity; >90% specificity; a significant improvement over assays currently available/in use in endemic zones).