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

Abstract

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

Human papillomavirus detection and typing using a nested-PCR-RFLP assay

“…[13][14][15][16] One of the main advantages of this primers combination is the possibility of determining the HPV type using different procedures: restriction fragment length polymorphism (RFLP) assay, sequencing, and micro-array technology. 10,[17][18][19][20] Nested-PCR techniques have proven to be more sensitive for the detection of HPV than PCR methods based on only one amplification reaction. 21,22 A nested-PCR method using MY09/11 and GP5+/6+ primers has been used in large epidemiological studies for the sensitive detection of HPV in cervical samples.…”