Desenvolvimento de metodologias para identificação molecular do HPV
Resumen
The Human Papiloma Virus (HPV) is a Sexually Transmitted Disease (STD) very
common in the world. It infects the human epithelium may persisting of asymptomatic
form or causing some neoplasia. Many studies report the association between HPV
and many kinds of cancer such as: lap utero, anus, penis, vagina and vulva. According
to INCA data for the year of 2016 are expected 16.340 new cases of lap utero cancer,
being the second most frequent case in the female population in Brazil. For the
recognition of the virus, there`s a lots of tracking methods, as morphological test (pap
test), that observes cytopathic effects caused by the virus on human cells, suggesting
the existence of infection, however this type of test presents low results and has shown
high taxes of false negative and positive results. To overcome this problems, countless
studies has shown the effect of molecular techniques utilization to increase the
sensibility and especially, getting recognize and genotyping the HPV virus. On this
recent studies, were tested distinct molecular techniques for typing the HPV virus, as
Conventional PCR followed by Sanger Sequencing , Real time PCR (SYBRGreen® e
Taqman ®) and Sequencing of New Generation. Altogether were collected 318 samples
pf cervix grated, and from this material were collected the DNA using an adapted
protocol (POWELL; GANNON, 2002). Using the conventional PCR technique followed
by Sanger Sequencing we obtained 65 positives samples for the HPV(21%), in 49
samples(75,3%) it was possible to identify the HPV type, in the other 16
samples(24,7%) it was not possible the identification, probably because the infection
was formed for two or more types of the virus. With the real time PCR technique using
SYBRGreen®, were accomplished an experimente with 30 samples, which was
possible to confirm the results in 28 of it, using Sanger Sequencing. In two samples
the results are not confirmed, being possible to positive the sample, showing high
sensibility of the real time PCR technique. The methodology of New Generation
Sequencing (NGS) it showed useful for HPV identification, being one of the first studies
published for routine use. And it has great prospects because besides HPV can identify
other microorganisms in the sample and quantifies them as well.