THINPREP HPV IN SITU HYBRIDIZATION WITH PNA PROBES

人乳头瘤病毒(HPV)与人类乳头瘤发生发展的关系 宫颈癌鼓励了对HPV检测方法的研究。这个 这项工作的目的是建立一种原位杂交方法，用于检测 薄片中HPV致病类型的检测 感染宫颈。ThinPrep技术是一种基于液体的过滤 一种允许对宫颈细胞进行清洗和单层的方法 幻灯片。这种清洗过程减少了血液、粘液和非诊断性疾病 产生用于宫颈筛查的高度可辨别的样本的碎片。瘦的 制备载玻片提高了检测异常细胞的灵敏度， 并减少了SBLB宫颈标本的数量(令人满意，但 受)地位的限制。在第一阶段，我们将开发一种原位HPV检测方法， 利用ThinPrep幻灯片、肽核酸探针和高度 灵敏的酪胺信号放大系统。在第二阶段，这项检测 将在细胞消化方法方面进行优化，细胞 固定，杂交严格条件，染色原检测，阻断 和洗涤剂。此后，这一检测方法将用于临床 为建立其在区分哪些女性应该是 积极治疗；即需要随访阴道镜或活组织检查。 第三阶段将专注于产品开发和临床试验。 建议的商业应用： 人乳头瘤病毒市场预计在未来五年内达到6000万美元 几年。一种能够进行分类的原位方法的商业化 有不同宫颈异常的患者将是第一个重要的 采取步骤实施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.