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High quality DNA from human papillomavirus (HPV) for PCR/RFLPs

2005; Instituto de Tecnologia do Paraná (Tecpar); Volume: 48; Issue: 1 Linguagem: Inglês

10.1590/s1516-89132005000100006

ISSN

1678-4324

Autores

Denise Wanderlei-Silva, Mariana Nobre, Rosa Silva Gonzaga, Luciana Silva Viana, Eduardo Ramalho Neto,

Tópico(s)

Molecular Biology Techniques and Applications

Resumo

The analysis of DNA in clinical samples for a secure diagnostic has become indispensable nowadays. Techniques approaching isolation of high molecular weigth DNA of HPV could lead to efficient amplification and early clinical diagnosis of the virus DNA by PCR (polymerase chain reaction). We describe a fast, non-toxical, efficient and cheap method for DNA isolation of human papilloma virus (HPV) from cervical smears using guanidine (DNAzol solution). A 450 bp DNA band correponding to the late region (L1) of the virus genome was detected by PCR, showing that the DNAzol extraction soluction generated a good viral DNA yield. The electrophoretic pattern after digestion with restriction endonucleases (RFLPs/PCR) revealed the predominance of HPV-16 and HPV-33 in the samples from the State of Alagoas, Brazil.

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