Lateral Flow Assay for Oncogenic Strains of HPV

HPV 致癌菌株的侧向层析检测

• 批准号: 6689043
• 负责人: LEONARD S BAZAR
• 金额: $ 10.7万
• 依托单位: TREVIGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2003-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6689043
• 关键词: 2,4 dinitrophenol; biotin; bromodeoxyuridine; cardenolides; cell line; clinical research; enzyme activity; fluoresceins; human papillomavirus; isothiocyanates; nucleic acid probes; nucleic acid quantitation /detection; oligonucleotides; oncogenic virus; phosphoester ligase; technology /technique development

DESCRIPTION (provided by applicant): Trevigen/TreviMed has developed the Mismatch Identification DNA Analysis System (MIDAS) that relies on SNIPase (genetically-optimized DNA mismatch repair enzymes) for the detection of pathogen DNA without the need for PCR amplification. The purpose of the research outlined in this Phase I proposal is to adapt MIDAS to a lateral-flow dipstick platform. To this end, we propose the following specific aims: (1) to design and test immobilized MIDAS probes to high-risk oncogenic strains of human papilloma virus (HPV strains 16 and 18) that incorporate biotin and various haptens such as dinitrophenyl (DNP), digoxigenin (DIG), fluorescein isothiocyanate (FITC), or bromodeoxyuridine (BrdU). In the presence of HPV target DNA and SNIPase, cleavage of the probe occurs, leading to the formation of a colored band on a lateral-flow dipstick. The MIDAS probes will be optimized initially with single-stranded oligonucleotides as targets, corresponding to HPV-16 and HPV-18 DNA sequences; (2) to optimize Dipstick-MIDAS for detecting the presence of cloned HPV-16 and HPV-18 DNA, and genomic DNA derived from HPV-16 and HPV-18-positive cell lines. These efforts will culminate in a rapid, sensitive, multiplexed, cost-effective, and accurate assay for oncogenic strains of HPV that cause cervical cancer.

描述（由申请人提供）： Trevigen/TreviMed开发了错配鉴定DNA分析系统（MIDAS），该系统依赖于SNIPase（遗传优化的DNA错配修复酶）来检测病原体DNA，而无需PCR扩增。本第一阶段提案中概述的研究目的是使MIDAS适应侧流量油尺平台。为此，我们提出了以下具体目标：（1）设计和测试固定化MIDAS探针的高风险致癌株的人乳头瘤病毒（HPV株16和18），纳入生物素和各种半抗原，如二硝基苯（DNP），地高辛（DIG），异硫氰酸荧光素（FITC），或溴脱氧尿苷（BrdU）。在HPV靶DNA和SNIPase存在的情况下，探针发生切割，导致在侧流试纸上形成有色条带。MIDAS探针最初将以单链寡核苷酸作为靶标进行优化，对应于HPV-16和HPV-18 DNA序列;（2）优化用于检测克隆HPV-16和HPV-18 DNA以及源自HPV-16和HPV-18阳性细胞系的基因组DNA的存在的Dipstick-MIDAS。这些努力将最终导致一种快速，敏感，多重，具有成本效益和准确的检测方法，用于检测导致宫颈癌的HPV致癌株。