Adams, David S.
Specific high-risk types of human papillomavirus (HPV) have been shown to cause cervical carcinomas. The purpose of this MQP was to develop type-specific assays for detecting specific HPV types in preserved pap smear samples. Three kinds of assays were tested: 1) immunofluorescence using type-specific antibodies, 2) RT-PCR to assay episomal (low risk) versus integrated (high-risk) HPV genomes, and 3) genomic PCR using high-risk type-specific primers while simultaneously inhibiting low-risk HPV amplification using PNA probes. Our data indicates that 1) immunofluorescence staining of cancerous cell lines was more effective than specimen cells, 2) RT-PCR of HPV mRNA was effective on HeLa cell lines, and more optimization is needed, 3) PCR amplification of high-risk HPV types could be induced with the devised primer cocktail while minimizing the amplification of low risk HPV types. 4) PNA probes specific for optimal binding to optimal HPV type 11 can inhibit amplification in PCR.
Worcester Polytechnic Institute
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Biology and Biotechnology