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The development of methods that directly detect HPV DNA has greatly changed the field of cervical cancer testing. Current guidelines include standard Papanicolaou (Pap) test cytology in conjunction with newer HPV DNA diagnostic tests. The two indications for HPV DNA testing are Pap equivocal results (ASC-US triage) and primary cervical cancer screening in women older than 30 years of age. Primary HPV screening of women older than 30 is 20% more sensitive than Pap testing, and though less specific, HPV-positive women with negative or borderline cytology can be monitored by repeat testing, thereby reducing total colposcopy referrals while maintaining improved detection of precancerous lesions. Because of the low cancer risk and unknown natural history of HPV infection in men, HPV testing of men in general is not recommended; however, screening by anal cytology and anoscopy may be beneficial among the high-risk group of men who may become infected with anal HPV.

Testing for HPV relies exclusively on molecular biology techniques using nucleic acid probes, because HPVs cannot be cultured in vitro. There is a single diagnostic assay that is approved by the US FDA for the specific detection of high-risk HPV infection. The hc2 High-Risk HPV DNA Test (Digene Diagnostics, Gaithersburg) is designed to collectively detect one or more of 13 HPV genotypes (16/18/31/33/35/39/45/51/52/56/58/59/68) having confirmed association with cervical cancer development. Digene also offers the hc2 HPV DNA Test, which differentiates between low risk types 6/11/42/43/44 and high risk types. The HC2 assays utilize cocktails of specific RNA probes that are directed toward target HPV DNA sequences. An antibody to DNA-RNA hybrids is used for both capture and detection, which uses labeled antibody for signal amplification in a microplate chemiluminescence detection system. Cervical
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