ASSAY FOR SIMULTANEOUS DETECTION OF MULTIPLE HPV TYPES

本项目的目的是开发一种快速、高度敏感的检测方法。 同时检测多个人的原位杂交 人乳头瘤病毒(HPV)型特异性DNA探针在单个 样本。在第一阶段，我们将使用一个由两个 已知HPV水平的特征良好的宫颈癌细胞系 16型和18型DNA。生物素和DNP标记人乳头瘤病毒16型的杂交 和HPV18型DNA探针将通过荧光显微镜可视化 在适当的荧光标记抗体与 DNA探针。各种荧光标记和抗体制剂将 接受测试。抗体封闭、结合和洗涤步骤将 最优化。提供最大信号强度的检测系统 在接下来的第二阶段中，将使用特异性和最小背景 在这些研究中，他们的检测方法对识别高和低的有效性 将对活检样本中的风险HPV类型进行评估。这些研究将 结果是一种简单的、临床相关的HPV鉴定方法 能够为大量的关键信息提供 最小的工作量、时间和样品材料。

The aim of this project is to develop a rapid, highly sensitive assay for simultaneously detecting the in situ hybridization of multiple human papilloma virus (HPV) type-specific DNA probes to the HPV DNA in a single specimen. In Phase I, we will be using a model system consisting of two well-characterized cervical caricinoma cell lines with known levels of HPV type 16 and 18 DNA. Hybridization of biotin- and DNP-labeled HPV type 16 and HPV type 18 DNA probes will be visualized by fluorescence microscopy following binding of the appropriate fluorescently labeled antibodies to the DNA probes. Various fluorescent labels and antibody preparations will be tested. Antibody blocking, binding, and washing steps will be optimized. The detection system which provides maximal signal intensity and specificity and minimal background will be used in subsequent Phase II studies in which the efficacy of their assay for identifying high and low risk HPV types in biopsy specimens will be assessed. These studies will result in a simple, clinically relevant HPV identification assay which is capable of supplying a considerable amount of critical information with a minimum of effort, time and sample material.