ASSAY FOR SIMULTANEOUS DETECTION OF MULTIPLE HPV TYPES

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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.