Development of consensus primers for amplification of human papillomavirus DNA.
|Title:||Development of consensus primers for amplification of human papillomavirus DNA.|
|Abstract:||The detection of human papillomavirus (HPV) sequences associated with anogenital tract infections is problematic due to the abundant diversity of HPV types found in the genital mucosa, the apparent geographic variation of HPV types, the prevalence of certain types in ethnic groups and the presence of yet unknown HPV types in anogenital samples. The objective of this work was to develop a technique that would allow for the detection of all HPVs in a variety of clinical samples: diseased tissues, latent infections, and samples with limited amount of material. Because of its promising potential, the polymerase chain reaction (PCR) was selected as the method of choice. To fulfill the objective of amplifying all HPV sequences, primers were designed from regions of homology identified in the E1 ORF from a selected group of HPV types. The primers were assayed with a variety of HPV types and their utility evaluated in clinical samples. After amplification, Southern blot hybridization was performed for accuracy of typing as well as to increase the sensitivity of the assay. PCR using E1 ORF consensus primers proved to be a successful tool in the detection of HPV DNA sequences in clinical samples. The primers are capable of amplifying known and unknown HPV types in diseased tissues and tissues latently infected with HPV. Their application allowed for the identification of potential new HPV sequences.|
|Collection||Thèses, 1910 - 2010 // Theses, 1910 - 2010|