The Studying Of Building The HPV DNA Detection Microarray With Technology Of Reverse Dot Blot In Haikou Area

Human papillomavirus (HPV) is an common oncogenic virus belongs to papovaviridae and papillomavirus. There are more than200types of HPV with different carcinogenic ability, according to their DNA sequence data.This thesis is in order to build the HPV DNA detection microarray with technology of reverse dot blot. Choosing13different HPV genotypes on the basis of the study of HPV molecular epidemiology in Haikou area to design specific primers and probes and create specific fragment clone library. The types include of HPV6,11,16,18,31,33,42,43,52,53,56,58and66.Among1045samples,436cases were infected by HPV. The positive rate was41.72%(436/1045). The cases of single infection, double infection and multiple infection were337,65and34. The constituent ratio were77.29%,14.91%and7.80%respectively. The proportion of high-risk HPV genotype was74.78%(252/337) in single infection. The most frequently detected HPV genotypes were HPV16, HPV58, HPV43, HPV52and HPV6with ratio of22.25%,17.89%,16.74%,16.06%and8.94%respectively. The infection rate of the three age groups(under the age of30,30～40and above40years old)were30.05%,27.06%and24.54%respectively.There were both no statistically significant between under30years of age and30-40age,under the age of30and above40years old(X2=0.8095, P>0.05; X2=3.0571, P>0.05).We successfully established the specific small fragment clones library of thirteen HPV types. Comparing with Genbank data, HPV16,31and53have four different sites respectively, other sequences are same. But the result of the phylogenetic tree show HPV16and HPV53have close genetic relationship,because of their sequences are in the same branch comparing with their own comparison. In other word,HPV31may be a new type but need much more evidence. The hospital laboratory detection result of sample p11091506was been infected by HPV39.Considering the infection rate of HPV39was1.15%in Haikou area,Our microarrays have no its hybridization sites.So our result without it. But we can’t completely ignored it with the changes in epidemiology.The other samples hybridize successfully. So we builded the HPV DNA genotyping microarray with technology of reverse dot blot successfully.