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）是白人和少数民族之间最大的健康差距之一。在美国，对于每一个TB感染的白色人，估计有9个非洲裔美国人、8个拉丁美洲人、6个美洲原住民、23个亚洲人和21个夏威夷原住民/太平洋岛民患有这种疾病[2]。与这种差异相结合的是结核病耐药突变的流行，这些突变具有相关的1000个多态性，跨越36个基因、两个启动子区和一个核糖体RNA编码区[3]。目前的方法主要适用于富裕的医疗保健社区，利用微生物培养，这需要复杂的实验室和数周才能确定结果。社会经济地位低下的少数群体难以与医生沟通和/或跟进，这可能导致缺乏适当的治疗。一种低成本、简单、快速的护理点（POC）检测方法可以检测结核病及其对一线药物（异烟肼、利福平、乙胺丁醇和吡嗪酰胺）的耐药性，这将改善这些少数民族社区的结核病诊断。然而，目前的技术缺乏实现该目标的灵敏度、特异性和/或多路复用能力。 因此，我们建议开发一种POC装置，该装置提供培养方法的灵敏度、核酸方法的特异性和突变的广泛覆盖。为了实现这一目标，我们将推进我们的TruArray平台（半球形多孔凝胶滴微阵列），用于使用芯片PCR和我们现有的MDR-TB PCR微阵列生物芯片进行结核病诊断。 在第一阶段，我们证明了我们的样品制备方法，我们的PCR微阵列生物芯片和原型POC仪器的可行性。对于第2阶段，我们建议扩大我们的测试覆盖范围，以包括对所有一线药物产生耐药性的其他突变，将该测试整合到我们的POC仪器中，并将该系统转移到墨西哥华雷斯的Medicos Especializados。

DESCRIPTION (provided by applicant): Of all diseases, tuberculosis (TB) represents one of, if not, the greatest health disparity between whites and minorities [1]. 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 that detects TB and its resistance to first-line drugs (isoniazid, rifampin, ethambutol,and pyrazinamide) would improve TB diagnostics for these minority communities. However, current technologies lack sensitivity, specificity, and/or multiplexing capacity to achieve this goal. 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 advance our TruArray platform (hemispherical porous gel drop microarrays) for TB diagnostics using on-chip PCR and our existing MDR-TB PCR-microarray biochips. During Phase 1 we demonstrated feasibility of our sample preparation approach, our PCR-micorarray biochips, and prototype POC instrument. For Phase 2 we propose to expand the coverage of our test to include additional mutations that confer resistance to all first-line drugs, integrate this test onto our POC instrument, and translae this system to Laboratorios Medicos Especializados in Juarez, Mexico.