Human Papillomavirus Infection and Virus Genotyping in Uterine Cervix Cancer in Venezuela
Human Papillomavirus Infection and Virus Genotyping in Uterine Cervix Cancer in Venezuela
1. Hypothesis:

 If high risk genotypes of human papillomavirus (HPV) is the cause of cervical cancer
 worldwide, the genotypes associated with cervical cancer in Venezuela would be the same
 types found in other countries.

 2. Objectives:

 Primary objective:

 The objective of this investigation is to determine the presence and genotypes of HVP
 infection in cervical intraepithelial neoplasia grade 2/3 (CIN 2/3), and Stage I cervical
 epidermoid carcinoma and cervical adenocarcinoma.

 Specific objectives:

 To establish the presence of HPV in cervical cancer patients in Venezuela by geographic
 distribution.

 To establish which HPV types are linked with cervical cancer in Venezuela by geographic
 distribution.
In Venezuela, cervical cancer is the second leading cause of cancer mortality in women. In
 developed countries, although it is not the first cause of cancer mortality, occupies a
 remarkable place in the neoplastic pathology (Cannistra, 1996; Dey, 1998).

 Human Papilloma Virus (HPV) is involved in the development of cervical cancer, specially
 oncogenic types 16 and 18 (Koskela, 2000; Markowska, 1999).

 To date, it is still unknown which HPV types are more frequent in Venezuela. Several studies
 made in this country concluded that HPV 16 is the predominant type in cervical pathology
 (Graterol 2006, Correnti 1997, Muñoz 2003), but there has not been performed a HVP genotyping
 to detect all the types involved in patients with cervical cancer. Neither has been establish
 which HPV types are more frequently involved in genital warts in Venezuela.

 The goal of this investigation is perform a genotypification of HPV in cervical cancer and
 cervical intraepithelial neoplasia 2 and 3 in Venezuela to determine which types are more
 frequently related to this disease in our country.

 4. STUDY DESIGN:

 4.1. Collection of cervical samples:

 1. Previous ethics committee approval (Instituto Venezolano de Investigaciones Cientificas
 and Instituto de Oncología Luis Razetti), we are going to collect paraffin blocks of 300
 patients with histological diagnosis of CIN 2-3 or Stage I cervical cancer.

 The samples will be obtained from archives of the Pathology Units of the following medical
 centers in Venezuela: Instituto de Oncología Luis Razetti, Anatomía Patológica Universidad
 Central de Venezuela(Caracas), Hospital Oncológico Pérez Carreño (Valencia), Maracaibo,
 Barquisimeto, Ciudad Bolívar, Puerto La Cruz, Mérida and Barinas.

 Inclusion criteria:

 1. Paraffin blocks with CIN 2/3, and Stage I cervical cancer biopsies obtained between 2001
 and 2011.

 2. Properly blocks identification in order to obtain personal and clinical patient
 information.

 4.2. Molecular biology procedures:

 1. Histologically handling of tissue: previous identification of pathologic lesions as
 described in inclusion criteria by an expert pathologist (sample with lesion but without
 necrosis), the paraffin block will be cut into 5 to 7 µm slides with new blades and
 gloves for each cut. It will be obtain 1 slides that will be distributed as follows: one
 sample will be send to Laboratorio de Virología Molecular del Instituto Venezolano de
 Investigaciones Científicas; the remained sample will be kept for further evaluation if
 is necessary. Each slide will be placed in a proper container and will be identified by
 a serial number in order to preserve patient intimacy and avoid biased results.

 2. Deparaffinisation as described in Coombs 1999.

 3. Tissue digestion, DNA extraction and precipitation as described in Gravitt 2000.

 4. HPV genotyping will be detected by INNOLiPA system. The results will be correlated with
 clinical information.
Inclusion Criteria:

 paraffin blocks of 329 patients with histologic diagnosis of CIN 2, CIN 3 or Stage I
 cervical cancer. -

 Exclusion Criteria:

 -