DNA circuits for point-of-care diagnostics

描述（由申请人提供）：我们建议调整新的熵驱动的DNA电路，该电路最近才被开发出来以满足即时诊断的迫切需求。亚稳态DNA回路将通过与特定mRNA等位基因杂交而被激活，并将经历一系列线性或指数的链交换扩增反应，最终产生可产生易于读取的比色信号的脱氧核酶过氧化物酶。我们将使用一种新的优化技术，顺序注射分析，以找到反应条件，在各种样品和环境条件下提供强大的信噪比。我们将优化的DNA电路应用于检测M.从阿富汗获得的痰液样本中发现结核病。这些实验应该为开发一种单管比色法用于耐药M的即时诊断奠定基础。肺结核感染。 公共卫生相关性：结合新的分析技术，新的优化方法，并在该领域的实际应用的影响，不能低估为推进公共卫生监测和维护的选择。同样的DNA回路可以很容易地重新设计，用于检测其他mRNA和等位基因，因此在第一世界的临床环境中也应该有很大的用途。

DESCRIPTION (provided by applicant): We propose to adapt novel, entropy-driven DNA circuitry that has only recently been developed to pressing needs in point-of-care diagnostics. The metastable DNA circuits will be activated by hybridization to a specific mRNA allele, and will undergo a series of strand exchange amplification reactions, either linear or exponential, culminating in the production of a deoxyribozyme peroxidase that can generate an easy-to-read colorimetric signal. We will use a novel optimization technique, sequential injection analysis, to find reaction conditions that give robust signal-to-noise ratios under a variety of sample and environmental conditions. We will apply the optimized DNA circuitry to the detection of rifampicin resistant RNA polymerase alleles in M. tuberculosis in sputum samples obtained from Afghanistan. These experiments should set the stage for the development of a one-tube, colorimetric assay for point-of-care diagnosis of drug resistant M. tuberculosis infection. PUBLIC HEALTH RELEVANCE: The impact of marrying novel assay techniques, novel optimization methods, and realistic applications in the field cannot be underestimated for advancing options for public health monitoring and maintenance. The selfsame DNA circuitry can be readily redesigned for the detection of other mRNAs and alleles, and should therefore find great use in first world clinical settings, as well.