Multiplex Microarray for Tuberculosis

描述（由申请人提供）：在世界范围内，结核病造成的人类死亡人数超过任何其他传染病，其中大多数死亡发生在发展中国家，这些国家缺乏获得改进诊断工具的机会。控制结核流行的关键是在潜伏性疾病转化为活动性疾病之前对患者进行早期识别和治疗。该SBIR项目解决了对确定潜伏性和活动性结核病（TB）感染个体的护理点检测的重要需求。基于血清学的检测很容易适用于医疗点资源，然而，对于结核病，没有定义诊断抗原决定因素。在研究的数十种结核病相关抗原中，没有一种具有足够的免疫优势，可以可靠地用于基于经典免疫测定的诊断。在这个项目中，我们用一种蛋白质多重微阵列来解决这个问题，这种微阵列能够在一次实验中检测到在对TB抗原的体液反应中产生的大量不同的抗体。该测试的诊断能力将随着可分析抗原数量的增加而直接增加。能够观察数百种确定的抗原，具有更高的灵敏度和特异性，应该允许更可靠和准确的分析，可以检测感染个体并区分潜伏和活跃感染阶段。所提出的平台比ELISA格式的灵敏度高几个数量级。这种增加的敏感性与检测IgA类抗体的便捷性相结合（在免疫功能低下的个体中，IgA反应通常不如IgG反应受影响），将允许对其他合并症（如HIV感染）患者使用该测试。在第一阶段，我们将用有限数量的临床定义样本测试多重微阵列平台的可行性。二期试验的成功完成将产生一种改进的、快速的结核病感染检测和筛查方法。

DESCRIPTION (provided by applicant): Worldwide, TB causes more human deaths than any other infectious disease and most of these deaths occur in developing countries where access to improved diagnostic tools is lacking. The key to controlling the TB epidemic is the early identification and treatment of patients with latent disease before conversion to active disease. This SBIR project addresses the important need for a point of care assay for identifying individuals infected with latent and active tuberculosis (TB). Serological based assays are readily adaptable to point-of-care resources, however, for TB, no diagnostic antigen determinants have been defined. Of the dozens of TB- associated antigens investigated, none are sufficiently immunodominant to be reliable for a classical immunoassay based diagnostic. In this project, we address this problem with a protein multiplex microarray that is capable of detecting in a single experiment, a large number of diverse antibodies that are generated in the humoral response to TB antigens. The diagnostic power of the test will increase directly with the number of antigens that can be analyzed. The ability to look at hundreds of defined antigens, with increased sensitivity and specificity should allow a more robust and accurate analysis which can detect infected individuals and differentiate between latent and active stages of infection. The proposed platform is orders of magnitude more sensitive than ELISA formats. This increased sensitivity combined with the ease of detecting IgA class antibodies (IgA response is often less effected than the IgG response in immune-compromised individuals) will permit the use of the test for patients with other co-morbidities e.g. HIV infection. In Phase I, we will test the feasibility of the multiplex microarray platform with a limited number of clinically defined samples. The successful completion of Phase II will produce an improved, rapid, point of care detection and screening assay for TB infection. Identification of individuals with active tuberculosis (TB) is critical to public health care efforts to prevent the spread of TB. However, accurately identifying and treating individuals who have latent TB, is vital to the over-all control of this disease. Worldwide tuberculosis (TB) causes more human deaths than any other infectious disease. Improved TB diagnostic services are less effective in immune compromised individuals and such services are extremely limited in countries with the highest TB burden and morbidity. An affordable, accurate, accessible immunoassay that enables early identification of latent or active TB is crucial for infection control program success. The proposed multiplex microarray methodology is a practical, efficient system to check for specific, significant TB antibodies. This point-of-care multiplex microarray assay for TB can identify at the point-of-care, latent as well as, active TB infection and therefore drug therapy can begin sooner.