Molecular detection of high-risk human papillomavirus (HPV) among Thai women and genome characterization of HPV16 and HPV18 in cytological samples of uterine cervix

Abstract

In this study, author established primers for high-risk HPV detection by using nucleotide sequencing method. HPV16 and HPV18 are the major cause of cervical cancer worldwide therefore, the author established HPV16 and HPV18 primer for whole genome characterization. In addition, the author established primers for HPV screening by specific at E1, E6 and L1 gene. Based on screening data of HPV distribution in this study, multiplex realtime PCR probes and primers were established for frequently detected genotypes. Focusing on the objective, seven and nine samples which derived from HPV16 and HPV18 infected women who have different cytological data were characterized whole genome, respectively. Analysis data discovered two critical changes in HPV16, converting the E2-219P prototype to E2-219T in cervical cancer and the L2-269S prototype to L2-269D in CIN III; however, there were no critical changing in HPV18. Next, 243 different cytological samples were analyzed by using E1, E6 and L1 primers. Author found that E1 primer was high percentage of detection (79.7%). Moreover, detection by E1 primer can be clarified samples which in HCII grey zone. Furthermore, author set up probe and primer for the highest prevalence genotypes HPV16 and HPV31 which confirmed result by testing with 515 samples by using E1 primer. Then multiplex real time PCR method was developed for HPV16, HPV31 and some additional HPV18 which is more important genotype today. In conclusion, primer designations were established for HPV screening by using PCR and multiplex real time PCR for nucleotide sequencing study which may be concern with mechanism and progressive to cancer. The author hopes that primer and probe in this study can be used for HPV studying and useful for public health study in the future.