tailieunhanh - Báo cáo y học: " Detection and quantitation of HPV in genital and oral tissues and fluids by real time PCR"

Detection and quantitation of HPV in genital and oral tissues and fluids by real time PCR | Seaman et al. Virology Journal 2010 7 194 http content 7 1 194 VIROLOGY JOURNAL METHODOLOGY Open Access Detection and quantitation of HPV in genital and oral tissues and fluids by real time PCR William T Seaman1 Elizabeth Andrews2 5 Marion Couch1 Erna M Kojic6 Susan Cu-Uvin6 Joel Palefsky7 Allison M Deal4 Jennifer Webster-Cyriaque1 2 3 Abstract Background Human papillomaviruses HPVs remain a serious world health problem due to their association with anogenital oral cancers and warts. While over 100 HPV types have been identified a subset is associated with malignancy. HPV16 and 18 are the most prevalent oncogenic types while HPV6 and 11 are most commonly responsible for anogenital warts. While other quantitative PCR qPCR assays detect oncogenic HPV there is no single tube assay distinguishing the most frequent oncogenic types and the most common types found in warts. Results A Sybr Green-based qPCR assay was developed utilizing degenerate primers to the highly conserved HPV E1 theoretically detecting any HPV type. A single tube multiplex qPCR assay was also developed using type-specific primer pairs and TaqMan probes that allowed for detection and quantitation of HPV6 11 16 18. Each HPV type was detected over a range from 2 X 101 to 2 X 106copies reaction providing a reliable method of quantitating typespecific HPV in 140 anogenital cutaneous oral benign and malignant specimens. 35 oncogenic and low risk alpha genus HPV types were detected. Concordance was detected in previously typed specimens. Comparisons to the gold standard detected an overall sensitivity of 89 95 CI 77 - 96 and specificity of 90 95 CI 52 - 98 . Conclusion There was good agreement between the ability of the qPCR assays described here to identify HPV types in malignancies previously typed using standard methods. These novel qPCR assays will allow rapid detection and quantitation of HPVs to assess their role in viral pathogenesis. Background Papillomaviridae comprise a diverse .