Lou2013 - Abstract
High-risk human papillomavirus infections are causally related to cervical cancer development. Cervical cancer represents the most dramatic cancer health disparity of women in the world, with over 200,000 deaths annually, concentrated in poor, rural, and indigenous populations. Guatemala and Venezuela are illustrative of this disparity as cervical cancer is the predominant cause of cancer cases and deaths in women. To determine the HPV viral strains in invasive tumors, we initiated a prospective collection of invasive cervical cancer cases with tissue collected in RNAlater. DNA-based testing has been shown to be useful in identifying persistent carriers at risk for cervical cancer. To determine the prevalence of HPV in women over 30, cervical swabs were collected at the time of a Pap exam and preserved in 3ml of Scope mouthwash. A total of 351 cancer cases with an average age of 53 (22-81 years), and 395 swabs of non-cancer cases over age 30 were obtained. We amplified the HPV L1 gene from DNA purified from cancer tissues and cell lysates from cervical swab samples using BS GP5+ and GP6+ primers and nested PCR was used to confirm the undetermined samples by primer MY09/11 (1st run PCR) and BS GP5+ and GP6+ (2nd) run PCR. DNA sequencing of HPV from tumor tissue revealed detectable in HPV in 96 10 0.8-4.9 cancer is approximately 12 determination we bar-coded the same BS GP5+ and GP6+ primers and sequenced on the Ion Torrent PGM. An average of 8000 reads were obtained for each sample. Highly comparable data was obtained as from the conventional sequencing and mixtures of HPV types as well as polymorphisms within the isolates was observed. From the HPV16 and HPV18 samples, oncoprotein E6 and E7 RNA was quantitated to estimate viral load. In conclusion, vaccination against HPV16 and HPV18 can protect for up to 70 will be required for many years. A sensitive and low cost approach to cervical sampling, including Next-Gen sequencing has been developed that could be employed to reduce the burden of cervical cancer worldwide.
Lou, H.; Villagran, G.; Odey, U.; Sawitzke, J.; Wells, D.; Troyer, J.; Dyba, M.; Ruch, A.; Orozco, R.; Argueta, V.; Gharzouzi, E.; Alvirez, E.; Dean, M. HPV and Cervical Cancer in Guatemala and Venezuela-Low-cost screening with NextGen sequencing. In Accepted for American Society of Human Genetics (ASHG) Meeting, Boston, MA, 2013 Oct. 22-26; .