Padronização da técnica de reação em cadeia da polimerase para diagnóstico do Papilomavírus humano

Abstract

The genital infection by the human papillomavims (Human Papillomavirus-HPV) is the most common sexually transmitted disease (STD) in women and in men. There are mom than 200 different types of HPV identified, among whom more than 100 are completely genetically sequenced and more than 120 with partial sequencing. The Hpv am classified according to the ability to change in cellular regulation, being divided into groups of low, intermediate and high risk. The methods used in the detection of HPV DNA in injuries vary widely in their sensitivity and specificity. Objectives: This study aimed to standardize the technique polymerase chain reaction-PCR, in the laboratory of molecular biology in the Faculdade de Ciencias da Satide of the Universidade Federal da Grande Dourados for diagnosis of Papillomatosis. Methodology: from mucous samples from the samples of Applied Microbiology Laboratory of the Faculdade de Ciencias da Satide began the detection and extraction processes processes of extraction and detection of viruses by polymerase chain reaction. For the extraction of DNA were tested three different protocols, following popular protocols for DNAzoI Tm, TRIzol -114 and extraction MiniPrep. Was elected the technique the PCR with the primers, MY09/MY I land GH20/GH21. The PCR products were subjected to electrophoresis in 3% agarose gel and fragments were visualized under ultraviolet light (UV) in transluminator. Results: of the four samples analyzed, all amplified the gene control of human 13-globin. Thus, the analysis of the presence of HPV DNA was made on all samples, who underwent PCR and were positive.