CMOS-Integrated Fluorescence Biochip SBIR Phase II Grant Application

 描述(由申请人提供)：第二阶段项目，由硅谷初创公司InSilixa，Inc.提交，建议利用第一阶段的成就，开发一个互补的金属氧化物半导体(CMOS)生物芯片平台，用于诊断传染病。该项目解决了抗生素耐药细菌日益增长的全球威胁，这些细菌的出现和传播现在使抗生素选择决策复杂化，对预后产生负面影响，增加医疗保健成本，并加剧耐药性水平。为了应对这一危机，InSilixa将开发一个高度多元化的平台，该平台可以同时、快速和廉价地检测临床样本中的感染性病原体，并识别导致耐药性的突变。具体地说，本申请中提出的工作将需要设计、实施和实验验证核酸扩增测试(NAAT)方法，该方法使用专有的Hydra CMOS生物芯片检测平台。目标1将开发一种单室、闭管和高度多重的NAAT技术，该技术实时和并行地定量10‘S的多重聚合酶链式反应，并通过执行100’S的高分辨率熔融曲线分析实验来鉴定和表征所产生的扩增产物的序列。AIM 2将进一步开发CMOS生物芯片检测平台(Hydra-4K)。Aim 3将在结合上述技术的基础上开发一种全面的NAAT检测方法，用于检测结核分枝杆菌的耐药性。虽然最初的临床重点是结核病，但在该项目过程中获得的设计、开发和制造技术将使Hydra平台的所有其他应用(和产品)成为可能。其中包括有吸引力的商业机会，包括呼吸道感染、抗药性医院获得性感染、门诊尿路和咽部感染，以及艾滋病毒/艾滋病患者血液中艾滋病毒的识别、定量和药物敏感性基因分型。这一二期项目的成功完成将导致InSilixa CMOS生物芯片平台的完全特征化的工作原型，该原型可以同时识别感染剂和导致对大量抗生素产生耐药性的突变。

 DESCRIPTION (provided by applicant): Phase II project, submitted by InSilixa, Inc., a Silicon Valley start-up company, proposes to leverage Phase I accomplishments by developing a complementary metal-oxide- semiconductor (CMOS) biochip platform for the diagnosis of infectious diseases. This project addresses the growing global threat of antibiotic resistant bacteria whose emergence and dissemination now complicates antibiotic selection decisions, negatively affects prognosis, increases health care costs, and fuels increasing levels of resistance. To tackle this crisis, InSilixa will develop a highly multiplex platform that can simultaneously, rapidly and inexpensively detect an infectious agent in a clinical sample and identify the mutations that confer drug resistance. Specifically, the work proposed in this application will entail the design, implementation, and experimental validation of a nucleic acid amplification testing (NAAT) method that uses the proprietary HYDRA CMOS biochip detection platform. Aim 1 will develop a single-chamber, closed-tube, and highly multiplexed NAAT technology that quantifies, in real- time and in parallel, 10's of multiplex PCR reactions and identifies and characterizes the sequences of the generated amplicons by performing 100's of high-resolution melt curve analysis (MCA) experiments. Aim 2 will further develop the CMOS biochip detection platform (HYDRA-4K). And Aim 3 will develop a comprehensive NAAT assay based on combining the above technologies for detecting antimicrobial-resistance in M. tuberculosis. Though the initial clinical focus is tuberculosis, the design, development and manufacturing know-how gained during the course of this project will enable all other applications (and products) for the HYDRA platform. Included amongst these are attractive commercial opportunities including respiratory tract infections, drug resistant hospital acquired infections, outpatient urinary tract and pharyngeal infections and the identification, quantificatin and drug susceptibility genotyping of HIV in the blood of HIV/AIDS patients. The successful completion of this Phase II project will result in a fully characterized, working prototype of the InSilixa CMOS biochip platform that can simultaneously identify an infectious agent and the mutations that confer resistance to a large panel of antibiotics.