DNA recovery from Hybrid Capture II samples stored in specimen transport medium with denaturing reagent, for the detection of human papillomavirus by PCR

• Published in: Journal of virological methods 2005-06, Vol.126 (1), p.197-201
• Main Authors: Rabelo-Santos, Silvia H., Levi, José Eduardo, Derchain, Sophie F.M., Sarian, Luis Otávio Z., Zeferino, Luiz Carlos, Messias, Samara, Moraes, Denise Lima de, Campos, Elisabete A., Syrjänen, Kari Juhani
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Cervix Uteri - cytology; Cervix Uteri - virology; Colposcopy; DNA extraction; DNA Primers; DNA Probes, HPV; DNA, Viral - analysis; DNA, Viral - genetics; DNA, Viral - isolation & purification; Female; Fundamental and applied biological sciences. Psychology; Globins - genetics; HPV; Human papillomavirus; Humans; Hybrid Capture II; Microbiology; Papanicolaou Test; Papillomaviridae - genetics; Papillomaviridae - isolation & purification; Polymerase chain reaction; Polymerase Chain Reaction - methods; Quality Control; Reference Standards; Specimen Handling - methods; Specimen transporting medium; Techniques used in virology; Vaginal Smears; Virology
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2005.02.009

The purpose of this study was to examine the quality of DNA recovered for human papillomavirus (HPV) detection using polymerase chain reaction (PCR) in samples that had been collected for Hybrid Capture II (HCII), testing and stored in specimen transport medium (STM) with denaturing reagent at −20 °C for 18 months. Endocervical tissue was collected from 92 women for HCII assay using the Digene STM, and a Papanicolaou smear was carried out in all cases. Seven women had normal colposcopy results. The remaining 85 patients underwent colposcopy-directed biopsy or cervical conization for histological investigation. Of the 92 samples tested, 84 were HCII-positive and 8 were negative. Quality control for amplification was carried out with β-globin primers G73 and G74, and HPV was tested using PGMY09 and PGMY11. DNA was recovered from 83 of the 92 samples (90%). Among the 84 samples HCII-positive initially, HPV was detected by PCR in 56 (67%). PCR did not detect HPV DNA in the eight samples that were HCII-negative, although five of them were positive for β-globin. This paper describes a novel DNA extraction technique that may permit exact HPV typing in stored samples collected originally for HCII testing, making it possible to carry out retrospective investigations to retrieve information on specific HPV types in large HCII series.