TB Diagnostics at the Point of Care

描述(由申请人提供)：在所有疾病中，结核病(TB)是白人和少数族裔之间最大的健康差距之一[1]。具体地说，在美国，每一个感染结核病的白人中，估计有9名非洲裔美国人、8名拉丁裔、6名美洲原住民、23名亚洲人和21名夏威夷/太平洋岛民原住民患有这种疾病[2]。与此不同的是结核病耐药突变的流行，这些突变具有相关的1000个多态，跨越36个基因、两个启动子区域和一个核糖体RNA编码区[3]。目前的方法主要适用于富裕的医疗保健社区，使用微生物培养，这需要复杂的实验室和数周的时间才能确定结果。处于较低社会经济阶层的少数群体在通勤和/或向医生跟进方面遇到困难，可能导致得不到适当的治疗。一种低成本、简单、快速的医疗点(POC)测试可以扩展 向这些少数民族社区提供耐药结核病诊断。然而，目前的技术缺乏敏感性、特异性和/或多路复用能力。 因此，我们建议开发一种POC设备，它提供培养方法的敏感性、核酸方法的特异性和对突变的广泛覆盖。为了实现这一目标，我们将扩展我们现有的MDR-TB聚合酶链式反应-微阵列生物芯片。这些生物芯片由印刷的凝胶元件微阵列组成，已被证明可以用凝胶元件中固定的引物扩增靶标。以前的工作表明，至少有60个独立的反应可以同时扩增1000个，在某些情况下是100个基因组拷贝，而不需要分裂，从而稀释样本。 我们的团队包括墨西哥华雷斯的实验室医疗队。来自该设施的团队成员将对我们的结核分枝杆菌(MTB)样本纯化设备进行初步评估，该设备之前在不列颠哥伦比亚省疾病控制中心(BC-CDC)的手中被证明是成功的。此外，华雷斯团队将验证Akonni的MDR-TB聚合酶链式反应-微阵列生物芯片。与此同时，Akonni将扩大耐药结核病阵列的多路复用能力，开发一种裂解方法，并将MDR-TB分析转化为Akonni的POC原型设备。在第二阶段，基因分型能力将进一步扩大，POC设备将被转移到华雷斯诊所。这项拟议的测试预计是3美元的消耗品，在5000美元的仪器上操作。 (与公共卫生有关的声明)：在所有疾病中，结核病是白人和少数族裔之间健康差距最大的疾病之一。具体地说，在美国，每一个感染结核病的白人中，估计有9名非洲裔美国人，8名拉丁裔，6名美洲原住民，23名亚洲人，以及21名夏威夷/太平洋岛民原住民。拟议的项目是开发一种护理点设备，用于识别可广泛传播给少数群体的耐药结核病菌株。

DESCRIPTION (provided by applicant): Of all diseases, Tuberculosis (TB) represents one of, if not, the greatest health disparity between whites and minorities [1]. To be specific, for every TB-infected white person in the United States, there are an estimated 9 African-Americans, 8 Latinos, 6 Native Americans, 23 Asians, and 21 Native Hawaiian/Pacific Islanders with this disease [2]. Compounded with this disparity is the prevalence of drug-resistant mutations of TB, which have an associated 1000 polymorphisms that span 36 genes, two promoter regions, and one ribosomal RNA coding region [3]. Current methodologies, available primarily to affluent healthcare communities, utilize microbial cultures, which require sophisticated laboratories and weeks before a result can be determined. Difficulties for minorities in a low socioeconomic class to commute and/or follow up with their physicians can result in a lack of appropriate treatment. A low-cost simple and rapid point-of-care (POC) test could expand drug-resistant TB diagnostics to these minority communities. However, current technologies lack sensitivity, specificity, and/or multiplexing capacity. We, therefore, propose to develop a POC device that offers the sensitivity of culture methods, specificity of nucleic acid methods, and a broad coverage of mutations. To accomplish this, we will expand upon our existing MDR-TB PCR-Microarray Biochips. These biochips consist of printed gel-element microarrays that have been shown to amplify target with immobilized primers in the gel elements. Previous work showed that at least 60 independent reactions can simultaneously amplify 1000, and in some cases 100 genomic copies, without needing to split, and thus dilute, the sample. Our team includes the Laboratorios Medicos Especializados in Juarez, Mexico. Team members from this facility will initially evaluate our sample purification device for Mycobacterium tuberculosis (MTB), previously shown to be sucessful at the hands of the British Columbia Centre for Disease Control (BC-CDC). Additionally, the Juarez team will verify Akonni's MDR-TB PCR-Microarray Biochip. In parallel, Akonni will expand the multiplexing capacity of the drug-resistant TB arrays, develop a lysis method, and translate the MDR-TB assay to Akonni's POC prototype device. During Phase II, the genotyping capacity will be expanded further and the POC devices will be translated to the Juarez clinic. This proposed test is projected to be a $3 consumable, operated on a $5000 instrument. (PUBLIC HEALTH RELEVANCE STATEMENT): Of all diseases, Tuberculosis (TB) represents one of, if not, the greatest health disparity between whites and minorities. To be specific, for every TB-infected white person in the United States, there are an estimated 9 African-Americans, 8 Latinos, 6 Native Americans, 23 Asians, and 21 Native Hawaiian/Pacific Islanders with this disease. The proposed project is to develop a point-of-care device for identifying drug-resistant strains of Tuberculosis that can be widely disseminated to minority populations.