Developing a POC CMOS biochip for oral HPV detection and quantification

开发用于口腔 HPV 检测和定量的 POC CMOS 生物芯片

• 批准号: 8974166
• 负责人: Arjang Hassibi
• 金额: $ 22.48万
• 依托单位: INSILIXA, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8974166
• 关键词: Archives; Bedside Testings; Biological Assay; Biological Markers; CDKN2A gene; Cancer Etiology; Cells; Characteristics; Clinic; Clinical Research; Consensus Sequence; DNA; Data; Dentists; Detection; Development; Devices; Diagnostic; Diagnostic tests; Drug resistance; Early Diagnosis; Engineering; Generic Drugs; Genes; Genome; Genomic DNA; Goals; HPV-High Risk; Head and Neck Cancer; Hela Cells; Housing; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16; Human papillomavirus 6; Incidence; Infection; Laboratories; Larynx; Length; Lesion; Liquid substance; Low risk HPV; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Measures; Messenger RNA; Molecular; Morbidity - disease rate; Mutation; Mycobacterium tuberculosis; Oncogenic; Oral; Oral cavity; Performance; Persons; Pharyngeal structure; Phase; Plasmids; Process; Proxy; Research Personnel; Risk; Scientist; Screening for Oral Cancer; Semiconductors; Small Business Innovation Research Grant; Specimen; System; Technology; Testing; Therapeutic Effect; Time; Transcript; Tube; Viral Genome; Viral Load result; Virus; Virus Diseases; Work; base; biochip; cohort; cost effective; design; follow-up; intraepithelial; metal oxide; molecular marker; mortality; non-oncogenic; oral infection; oral tissue; point of care; point-of-care diagnostics; prognostic; public health relevance; repository; research study; sensor; tumor; viral DNA; viral detection

 DESCRIPTION (provided by applicant): Human Papilloma Virus (HPV) oral infection is responsible for an increasing percent of head and neck cancers. The risk of progression from infection to cancer is increased by persistent HPV infection, infection with high-risk HPV types, high viral load and increased abundance of HPV E6/E7 transcripts and host p16INK4A transcripts. Rapid cost-effective point- of-care (POC) tests that detect and quantify risk-associated biomarkers in oral biospecimens are needed for oral cancer screening, prognostic purposes and to follow-up persons treated for high-grade lesions. Hypothesis: We hypothesis that it is possible to use the InSilixa HYDRA-1K CMOS biochip to develop a multiplex assay that can detect, identify and quantify HPV high and low risk types in oral tissues and fluids. Preliminary Data: Co-investigators at UNC have developed a non POC multiplex qPCR assay that can detect, identify and quantify HPV 6, 11, 16 and 18 L1 gene type-specific sequences and HPV E1 gene consensus sequences common to all HPV types. This assay has been used in clinical studies of oral biosamples. InSilixa engineers have developed the 1,024 sensor pixel multiplex HYDRA-1K CMOS biochip and used this platform to identify 117 separate SNPs in the M. tuberculosis genome that confer drug resistance. Specific Aims: The proposed Phase I project seeks to develop a complementary metal-oxide-semiconductor (CMOS) platform, the InSilixa HYDRA-1K, as a POC molecular diagnostic device that can detect, type and quantify HPV DNA in oral biospecimens. In Specific Aim 1 we will design, fabricate, test and optimize HPV type-specific and generic capture probes attached to a dynamic microarray experimental setup and proxy for the InSilixa HYDRA-1K biochip. In Specific Aim 2 we will design, fabricate, test and optimize primers for an asymmetric quantitative multiplex PCR assay that amplifies regions of the HPV genome that contain HPV type-specific and consensus sequences. In Specific Aim 3 we will determine the performance specifications of a HPV dynamic microarray assay that integrates the capture probes from Specific Aim 1 with the asymmetric quantitative multiplex PCR assay from Specific Aim 2. Successful completion of these Specific Aims will provide the basis for a Phase II SBIR application that will migrate the HPV assay developed here from the dynamic microarray setup to the HYDRA-1K CMOS platform.

 描述（由适用提供）：人类乳头状瘤病毒（HPV）口腔感染导致头颈癌的百分比增加。通过持续的HPV感染，高风险HPV类型的感染，高病毒载荷以及HPV E6/E7转录本的抽象增加以及宿主P16INK4A转录物的增加，从感染到癌症的风险增加了。在口腔癌筛查，预后目的和对高级病变治疗的后续人员中，需要快速成本效益的护理测试（POC）测试，这些测试需要在口腔生物测量中检测和量化与风险相关的生物标志物。假设：我们假设可以使用Insilixa Hydra-1K CMOS生物芯片来开发一种可以检测，识别和量化口腔组织和烟道中HPV高和低风险类型的多重分析。初步数据：UNC的共同研究器开发了一种非POC多重QPCR测定法，该测定方法可以检测，识别和量化HPV 6、11、16和18 L1基因类型特异性序列和HPV E1基因共有序列，所有HPV类型常见。 Insilixa工程师已经开发了1,024个传感器像素多重氢-1K CMOS Biochip，并使用该平台在结核分枝杆菌基因组中识别了抗药性耐药性的117个单独的SNP。具体目的：拟议的I期项目旨在开发一个完整的金属氧化物 - 氧化 - 氧化氢（CMOS）平台，即Insilixa Hydra-1K，作为一种POC分子诊断设备，可以检测，类型和量化口服生物素中的HPV DNA。在特定的目标1中，我们将设计，制造，测试和优化与Insilixa Hydra-1K Biochip的动态微阵列实验设置相关的HPV类型特异性和通用捕获问题。在特定的目标2中，我们将设计，制造，测试和优化引物，用于不对称定量多重PCR测定法，该测定法，该测定法的HPV基因组的放大器区域包含HPV类型特异性和共识序列。在特定目标3中，我们将确定集成捕获问题的HPV动态微阵列测定的性能规范。从特定的目标1带有来自特定目标2的不对称定量多重PCR分析。这些特定目标的成功完成将为II期SBIR应用提供基础，该应用将迁移此处开发的HPV测定法，从动态微阵列设置到Hydra-1K CMOS平台。