title: Detection and quantitation of HPV in genital and oral tissues and fluids by real time PCR.
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 10(1) to 2 x 10(6)copies/reaction providing a reliable method of quantitating type-specific 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%).
  
  
  CONCLUSIONS
  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.
authors: Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, NC, USA.
authors: William T
authors: WT
authors: Seaman
authors: Elizabeth
authors: E
authors: Andrews
authors: Marion
authors: M
authors: Couch
authors: Erna M
authors: EM
authors: Kojic
authors: Susan
authors: S
authors: Cu-Uvin
authors: Joel
authors: J
authors: Palefsky
authors: Allison M
authors: AM
authors: Deal
authors: Jennifer
authors: J
authors: Webster-Cyriaque
mesh_terms: D052160:Benzothiazoles; D001826:Body Fluids / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D003959:Diamines; D005260:Female; D005835:Genitalia / Q000821:virology*; D006801:Humans; D008297:Male; D009055:Mouth / Q000821:virology*; D009930:Organic Chemicals / Q000378:metabolism; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D014764:Viral Proteins / Q000235:genetics; D014773:Virology / Q000379:methods*
chemical_list: D052160:Benzothiazoles; D017931:DNA Primers; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; D014764:Viral Proteins; C098022:SYBR Green I