Detection and Typing of the Human Papilloma Virus in Women with Cervical Cancer in Bogotá and Barranquilla. Technical and Diagnostic Procedures
Keywords:
HPV, DNA typing, uterine cervical neoplasms, molecular diagnostic techniques, Colombia.Abstract
Objective: To analyze the role played by technical and diagnostic techniques in the detection of diverse genotypes of the human papilloma virus (HPV) in cervical cancer samples.
Methods: Paraffin fixed tissue samples were taken from 268 cases of cervical cancer in Bogotá and Barranquilla. Histological diagnosis was verified through new cuts and sample quality was analyzed through amplification of the β-globin gene. In the case of β-globin negatives, additional cuts and amplification were carried out. HPV detection was achieved using GP5+/GP6+bio indicators directed towards the L1 region, and typing through EIA and RLB. Among GP5+/GP6+ negatives, specific PCR type was developed towards the E7 region of 14 types of high risk HPV.
Results: Out of the initial 268 samples, 20 (7.46%) had inadequate histological diagnosis. Initially β-globin negative samples came to 55/248, but 29 recuperated with the realization of a second PCR, resulting in 26 β-globin negatives (10.5%). HPV was detected in 210/222 appropriate samples through GP5+/GP6+ (94.6%), and in 7 additional samples (3.15%) through the indicators directed towards E7. No infection was detected in 2.25% of cases. Twenty-four types of HPV were found, the most prevalent being HPV-16 (50.9%), HPV-18 (12.7%), HPV-45 (8.8%), HPV-31 (6.5%), and HPV-58 (6.0%). Multiple infection occurred in 16.6% of cases.
Conclusions: The adequate processing and diagnosis of samples and the use of combined tests towards the L1 and E7 regions allows for the optimum appraisal of infection prevalence among paraffin fixed samples.
Author Biographies
Mónica Molano, Instituto Nacional de Cancerología
Grupo de Investigación en Biología del Cáncer, Instituto Nacional de Cancerología, Bogotá. Colombia.
Raúl Murillo, Instituto Nacional de Cancerología
Subdirección de Investigaciones, Instituto Nacional de Cancerología, Bogotá. Colombia.
Arlenne Cano, Instituto Nacional de Cancerología
Subdirección de Investigaciones, Instituto Nacional de Cancerología, Bogotá. Colombia.
Óscar Gamboa, Instituto Nacional de Cancerología
Subdirección de Investigaciones, Instituto Nacional de Cancerología, Bogotá. Colombia.
Jaime Ardila, Instituto Nacional de Cancerología
Subdirección de Investigaciones, Instituto Nacional de Cancerología, Bogotá. Colombia.
Juan Carlos Mejía, Instituto Nacional de Cancerología
Grupo de Patología, Instituto Nacional de Cancerología, Bogotá. Colombia.
Gilberto Martínez, Clínica del Country
Grupo de Ginecología, Clínica del Country, Bogotá. Colombia.
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