No abstract

Infection with a high-risk carcinogenic type of human papillomavirus (HPV) is necessary for the development of cervical cancer. The digene HC2 HPV Test (HC2) is an important screening tool but lacks genotyping capability. To address this issue , we developed an assay for the rapid genotyping of HPV in cervical specimens. The three steps of this assay include Hybrid Capture target enrichment , whole-genome amplification , and Luminex XMAP detection. The assay includes the simultaneous detection of two genomic regions from each of 17 high-risk and two low-risk HPV types most associated with disease. The assay performance was tested on HPV plasmids as well as clinical specimens. An analytical limit of detection of 100 copies or less was demonstrated for linear, circular, and integrated HPV DNA. This finding is at least 1 log lower than the HC2 assay limit of detection. There was no cross-reactivity among the HPV types up to 1,000,000 copies. There was also no substantial assay interference from substances in cervical specimens. Although the clinical performance of the assay was not formally tested, the assay had good agreement (Cohen's kappa equal to 0.72) with both a PCR-based HPV genotyping assay (n ‫؍‬ 131) and the HC2 assay (n ‫؍‬ 502) using representative cervical specimens. This assay may be easy to automate and could be applied for the detection of other targets in future studies. Infection with a carcinogenic type of human papillomavirus (HPV) is necessary for development of cervical cancer.1,2 There are 13 types of HPV (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68) confirmed to be of high or moderate risk for cervical cancer and four types (26, 73, 66 and 82) that are probable high-risk. Other HPV types are low-risk and may be associated with genital warts, such as HPV 6 and 11. Molecular-based screening for the most oncogenic types of HPV is being used more frequently to diagnose risk for cervical cancer because it is easier and more reliable than cytological screening. 3For example, the digene HC2 HPV DNA Test (HC2) is widely used for screening 4 because it has exceptional clinical performance for a cancer and high-grade disease end point (ie, negative predictive value). Molecular screening, however, is not designed to determine which HPV type is present in an infection. The type of HPV in an infection is important to monitor because the risk of cancer and high-grade lesions increases for those whose HPV infection type is the same over time, especially for HPV 16 and 18.