Quantitation of human papillomavirus type 16 E6 oncogene sequences by real-time or quantitative PCR with EvaGreen.
Quantitation of E6 oncogene sequences of the human papillomavirus type 16 by real-time or quantitative PCR (qPCR) is used to determine the viral load, which correlates with the degree of the cervical neoplastic lesions. In the presence of EvaGreen, a new DNA intercalating fluorochrome, we obtained consistent and reproducible qPCR amplification curves and thermal denaturation profiles identical to those of the authentic E6-HPV16 (human papillomavirus 16) genome from the amplification products derived from a construct carrying the E6-HPV16 oncogene. E6-HPV16 quantitation in the presence of EvaGreen, therefore, is reproducible and specific and may be used to determine HPV16 viral load.
División de Biologi a Molecular, Instituto Potosino de Investigación Cienti fica y Tecnológica, 78216 San Luis Potosi, Mexico.
Socorro
S
Hernández-Arteaga
Rubén
R
López-Revilla
D001483:Base Sequence; D005456:Fluorescent Dyes / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D007364:Intercalating Agents / Q000032:analysis / Q000737:chemistry; D008969:Molecular Sequence Data; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012097:Repressor Proteins / Q000235:genetics*; D013997:Time Factors
C068651:E6 protein, Human papillomavirus type 16; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D009856:Oncogene Proteins, Viral; D012097:Repressor Proteins