Development and validation of a real-time PCR assay specifically detecting human papillomavirus 52 using the Roche LightCycler® 480 system

The recently released Linear Array® human papillomavirus (HPV) genotyping test (Roche Diagnostics) provides a standardized method for simultaneous detection of up to 37 individual HPV types. This test offers a rapid approach for detecting and longitudinal monitoring of patients infected with high-risk (HR) HPV. However, it cannot rule out HPV52 infections in the presence of carcinogenic HPV genotypes 33, 35 and/or 58. As such, often only a non-definitive result of HPV52 presence can be reported. This study describes the development of a real-time PCR assay using an HPV52-specific hydrolysis probe in conjunction with the newly released Roche LightCycler® 480 system. HPV52 was readily detected among DNA extracts from samples previously identified with possible HPV52 by the linear array test. Specificity was analyzed using a panel of DNA extracts previously identified as containing single/multiple HPV types, with or without HPV52. This is a rapid and simple assay, which could be used as a supplementary test to the Linear Array, to verify the presence or absence of HPV52 among samples testing positive for either HPV33, 35, and/or 58. Laboratories already using the Linear Array HPV genotyping test could adopt this method once internally validated.