Detection and quantitation of HPV gene expression using real-time PCR.

Human papillomaviruses (HPVs) are known to be etiological agents of cervical cancer and have been found in 99.7% of women with high-grade (HG) cervical intraepithelial neoplasia (CIN) precancer. Testing of high-risk HPV (HR-HPV) has been proposed as a way of improving cervical screening, especially for women with low-grade (LG) Papanicolaou (Pap) smears. In this chapter, real-time quantitative polymerase chain reaction (PCR) methods that can be used to investigate the expression of HPV 16 early genes in HG or LG precancer are demonstrated. Detecting the expression of early HPV genes in conjunction with the Pap smear may improve the specificity of identifying LG precancers that are associated with high risk of progression.