Abstract
Detection of integrated HPV DNA in SiHa and CaSki cells has been demonstrated by in situ PCR (1,2,3) and more recently TSA (4). These reports focused on high sensitivity single copy HPV detection using chromogenic techniques. Here we utilize high resolution and high sensitivity fluorescent methods of detection to analyze the spatial relationship of HPV DNA with respect to subnuclear domains enriched in splicing-related proteins, shown here by immunofluorescent detection of the SC-35 antigen (5). In situhybridization was performed using conventional reagents and the resulting probe signal amplified using fluorescent labeled tyramides (6). In contrast to techniques such as in situ PCR which require specialized procedures and equipment, detection of HPV DNA in SiHa cells was rapidly and routinely achieved using no specialized equipment. In addition, high target resolution was maintained, in contrast to in situ PCR, which is plagued by signal dispersion and difficult localization (7).
Published Version
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