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（由申请人提供）：人乳头状瘤病毒（HPV）口腔感染是导致头颈部癌症百分比增加的原因。持续性HPV感染、高危型HPV感染、高病毒载量和HPV E6/E7转录物和宿主p16 INK 4A转录物丰度增加会增加从感染进展为癌症的风险。检测和量化口腔生物样本中风险相关生物标志物的快速成本效益的床旁（POC）测试对于口腔癌筛查、预后目的和随访治疗高级别病变的人是需要的。假设：我们假设有可能使用InSilixa HYBRID-1 K CMOS生物芯片开发一种多重检测方法，可以检测，识别和定量口腔组织和液体中的HPV高风险和低风险类型。初步数据：共同研究者在ESTA开发了一种非POC多重qPCR检测，可以检测，鉴定和定量HPV 6，11，16和18 L1基因类型特异性序列和HPV E1基因共有序列常见的所有HPV类型。该测定法已用于口腔生物样品的临床研究。InSilixa的工程师开发了1,024个传感器像素的多路复用HYBRID-1 K CMOS生物芯片，并使用该平台识别了M.结核病基因组赋予耐药性。具体目标：拟议的第一阶段项目旨在开发一种互补金属氧化物半导体（CMOS）平台，InSilixa HYBRID-1 K，作为一种POC分子诊断设备，可以检测，分型和定量口腔生物标本中的HPV DNA。在具体目标1中，我们将设计，制造，测试和优化HPV类型特异性和通用捕获探针连接到动态微阵列实验装置和InSilixa HYBRIDGE-1 K生物芯片的代理。在特定目标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检测，该检测扩增含有HPV类型特异性和共有序列的HPV基因组区域。在Specific Aim 3中，我们将确定HPV动态微阵列检测的性能质量标准，该检测将Specific Aim 1的捕获探针与Specific Aim 2的不对称定量多重PCR检测相结合。这些特定目标的成功完成将为第二阶段SBIR应用提供基础，该应用将把此处开发的HPV检测从动态微阵列设置迁移到HYPERTIC-1 K CMOS平台。