THINPREP HPV IN SITU HYBRIDIZATION WITH PNA PROBES

人乳头瘤病毒（HPV）与宫颈癌发生的关系 宫颈癌鼓励研究HPV检测方法。的 这项工作的目的是开发一种原位杂交检测方法， 在Thin Prep载玻片上检测广泛的致病性HPV类型 感染子宫颈ThinPrep技术是一种基于液体的过滤技术 方法，允许清洁和单层的子宫颈细胞， 公里.这种清洁过程减少了血液，粘液和非诊断性 产生高度可辨别的样本用于子宫颈筛查。薄 制备载玻片提高了检测异常细胞的灵敏度， 并减少了宫颈标本的数量与SBLB（满意，但 受限于）状态。在第一阶段，我们将开发一种原位HPV检测方法， 利用ThinPrep载玻片、肽核酸探针和高度 灵敏的酪胺信号放大系统。在第二阶段，本试验 将在细胞消化、细胞培养和细胞培养方法方面进行优化。 固定，杂交严格条件，色原检测，阻断 和洗涤试剂。此后，该测定将用于临床试验。 这是为了确定其在区分哪些妇女应该被 积极治疗;即，需要后续阴道镜检查或活检。 第三阶段将专注于产品开发和临床试验。 拟定商业应用： HPV市场预计将在未来五年内达到6000万美元 年。能够进行分类的原位方法的商业化 不同宫颈异常的患者将是重要的第一个 采取更具成本效益和效率的方法， 检测和治疗宫颈癌高危妇女。

The association between human papillomavirus (HPV) and the development of cervical cancer has encouraged research in methods for HPV testing. The objective of this effort is to develop an in situ hybridization assay for detecting on Thin Prep slides the wide range of pathogenic HPV types infecting the cervix. ThinPrep technology is a liquid based filtration method that allows the cleansing and monolayering of cervical cells on slides. This cleansing process reduces blood, mucus, and nondiagnostic debris yielding highly discernible specimens for cervical screening. Thin Prep slides have improved the sensitivity in detection of abnormal cells, and reduced the number of cervical specimens with SBLB (Satisfactory But Limited By) status. In Phase I, we will develop an in situ HPV assay that utilizes ThinPrep slides, peptide nucleic acid probes, and the highly sensitive tyramide signal amplification system. In Phase II, this assay will be optimized with respect to methods of cell digestion, cell fixation, hybridization stringency conditions, chromogen detection, block and wash reagents. Thereafter, this assay will be used in a clinical setting to establish its potential in discriminating which women should be treated aggressively; i.e., requiring follow up colposcopy or biopsy. Phase III will focus on product development and clinical trials. PROPOSED COMMERCIAL APPLICATION: The HPV market is anticipated to reach $60,000,000 in the next five years.The commercialization of an in situ method capable of triaging patients with different cervical abnormalities will be an important first step in implementing a more cost effective and efficient approach for detecting and treating women at high risk for cervical cancer.