Projects
Name
Infection and risk factors of Human Papillomavirus (HPV) in women attending cervical screening in the Southeast of Brazil.
University
State University of Sao Paulo (UNESP)
Domain
Pathology
Departement
Department of Pathology Botucatu Medical School - FMB Sao Paulo State University ? UNESP
Head
Maria Aparecida Marchesan Rodrigues, MD, PhD
Tutor
Marcia Guimaraes da Silva, PhD/ Gabriel Vitor da Silva Pinto, MSc.
Languages
English, Spanish, Portuguese
Duration
4 weeks
Availability
Cities/Months Jan Feb Mar Apr May Jun Jul Augt Sep Oct Nov Dec
No No No No No No Yes Yes Yes Yes No No
Type of Research Project
- Clinical Project with Laboratory work
What is the background of the project?
Genital infections with oncogenic types of Human Papillomavirus (HPV) are very common among sexually active women, but only a minority of them will have a persistent HPV infection and develop cervical cancer. High-risk HPV infections are also associated with other anogenital diseases, such as 85% of anal cancers and 50% of the cancers ofvulva, vagina and penis. While high-risk HPV types have been recognized as the major causative agents for both malignancies and precursor lesions, other factors may also be involved and seem to act along with HPV infection, such as other sexually transmitted infections (STI), smoking or host immunologic response. Several behaviors, such as oral contraceptive use, sexual activities, tobacco and alcohol consumption, may influence the risk of initial viral infection and subsequent clearance of the virus. Considering the economic importance and the health impact of cervical cancer in developing countries, there is a special need for prospective studies regarding HPV prevalence and its risk factors.
What is the aim of the project?
The goals of this study were to determine the HPV infection rate in reproductive-aged women and to identify some risk factors associated with this infection.
What techniques and methods are used?
DNA from cervical samples will be extracted using the AmpliLute Liquid Media Extraction Kit (Roche Molecular Systems, Inc.) according to manufacturer's instructions. The HPV genotyping will be performed using the Linear Array Genotyping Test (Roche Molecular Systems, Inc.) that is able to identify 37 HPV genotypes. Reactions will be amplified in a PerkinElmer TC9600 Thermal Cycler. A positive control and a negative control for HPV detection, provided by Roche Molecular Systems, will be performed in each assay. The human globin target will be co-amplified to determine the sample adequacy. The evaluation of CT will be done with specific primers for PCR: CT1 (5TAG TAA CTG CCA CTT CAT CA 3), CT2 (5 TTC CCC TTG TAA TTC GTT GC 3), PL6.1 (5 AGA GTA CAT CGG TCA ACG A 3) and PL6.2 (5 TCA CAG CGG TTG CTC GAA GCA 3). PCR will be performed using the Go Taq Green master mix (Promega Corporation, USA). Amplification parameters will consist of40 cycles of 1 min at 95oC, 1 min at 55oC and 1 min and 30s at 72oC.
What is the role of the student?
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
DNA from cervical samples will be extracted using the AmpliLute Liquid Media Extraction Kit (Roche Molecular Systems, Inc.) according to manufacturer's instructions. The HPV genotyping will be performed using the Linear Array Genotyping Test (Roche Molecular Systems, Inc.) that is able to identify 37 HPV genotypes. Reactions will be amplified in a PerkinElmer TC9600 Thermal Cycler. A positive control and a negative control for HPV detection, provided by Roche Molecular Systems, will be performed in each assay. The human globin target will be co-amplified to determine the sample adequacy. The evaluation of CT will be done with specific primers for PCR: CT1 (5TAG TAA CTG CCA CTT CAT CA 3), CT2 (5 TTC CCC TTG TAA TTC GTT GC 3), PL6.1 (5 AGA GTA CAT CGG TCA ACG A 3) and PL6.2 (5 TCA CAG CGG TTG CTC GAA GCA 3). PCR will be performed using the Go Taq Green master mix (Promega Corporation, USA). Amplification parameters will consist of40 cycles of 1 min at 95oC, 1 min at 55oC and 1 min and 30s at 72oC.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
1
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
It is required that the student has some skills in basic laboratory techniques such as buffers preparation, pipetting and laboratory safety measures.
Are there any legal limitations in the student’s involvement
No
Hours
8
Type of students accepted
This project accepts:
- Medical students
Articles
- Baseman JG; Koutsky LA. The epidemiology of human papillomavirus infections. J ClinVirol 2005; 1:S16-24.
- Rosa MI; Fachel JM; Rosa DD; et al. Persistence and clearance of human papillomavirus infection: a prospective cohort study. Am J ObstetGynecol 2008;199(6):617.e1-7.