title: Efficient treatment of paraffin-embedded cervical tissue for HPV DNA testing by HC-II and PCR assays.
abstract: OBJECTIVE
  High-risk human papillomaviruses (HR-HPVs) are the primary cause of cervical cancer. In order to meet with clinical requirements, a direct capture test with signal amplification (HC-II), able to detect the 13 prevalent HR-HPVs, has been developed and validated for cytological specimens.
  
  
  METHODS
  the use of HC-II assay with formaldehyde-fixed paraffin-embedded cervical biopsies, for retrospective studies or to support histological findings, was investigated by analysing three different sample treatments. The efficacy of this test was compared with a reference PCR-ELISA, using MY09/11 and GP5+/6+ consensus primers and the use of a single extraction method for both HC-II and PCR-ELISA assays was validated.
  
  
  RESULTS
  protease treatment of dewaxed biopsy sections allowed an optimal performance of HC-II and has also been validated for PCR-ELISA. Overall, on the analysis of 50 cervical samples HC-II and PCR-ELISA assays showed a high concordance (K=0.80). Compared with PCR-ELISA, the HC-II had a sensitivity of 86.4% and a specificity of 92.9%. The results showed that short amplimers are necessary for a sensitive PCR-ELISA from formaldehyde-fixed paraffin-embedded biopsies, while HC-II showed a relatively low sensitivity for HPV18 within the HR probe pool.
  
  
  CONCLUSIONS
  HC-II can be a valid tool for the diagnosis of HPV infection in biopsy material. The possibility to use the same specimen preparation material for both HC-II and PCR-ELISA allows HC-II positive specimens to be further processed by PCR-ELISA if specific genotyping is needed.
authors.affiliation: Department of Clinical and Experimental Medicine, Section of Microbiology, University of Bologna, Via Massarenti 9, 40138 Bologna, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Cricca
authors.forename: F
authors.initials: F
authors.lastname: Bonvicini
authors.forename: S
authors.initials: S
authors.lastname: Venturoli
authors.forename: S
authors.initials: S
authors.lastname: Ambretti
authors.forename: G
authors.initials: G
authors.lastname: Gallinella
authors.forename: G
authors.initials: G
authors.lastname: Gentilomi
authors.forename: M
authors.initials: M
authors.lastname: Musiani
authors.forename: M
authors.initials: M
authors.lastname: Zerbini
mesh_terms: D001706:Biopsy; D002584:Cervix Uteri / Q000473:pathology / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D010450:Endopeptidases / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D027383:Papillomaviridae / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000821:virology; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014021:Tissue Preservation / Q000379:methods*
chemical_list: D004279:DNA, Viral; D010450:Endopeptidases