title: Comparative evaluation of the GP5+/6+, MY09/11 and PGMY09/11 primer sets for HPV detection by PCR in oral squamous cell carcinomas.
abstract: The aim of this study was to evaluate the use of GP5+/6+, MY09/11 and PGMY09/11 primer sets for the detection of human papillomavirus (HPV) DNA by single step polymerase chain reaction (PCR) and nested PCR in formalin-fixed and paraffin-embedded (FFPE) tissues from oral squamous cell carcinomas (OSCCs). DNA extracted from FFPE tissues were tested for amplification of the human beta globin gene with PCO3/4 primers. Positive samples for this gene were tested for HPV DNA using single step PCR with GP5+/6+, MY09/11 and PGMY09/11 primer sets. All negative samples at single step PCR with MY09/11 and PGMY09/11 were subjected to a further PCR with GP5+/6+ primers using the non-amplified product in the previously reactions (nested PCR) as samples. Among 26 samples, 23 were positive for the human beta globin gene and were considered viable for HPV DNA detection by PCR. Single step PCR with GP5+/6+ and MY09/11 primers and MY/GP+ nested PCR did not amplify HPV DNA in any samples. PGMY09/11 primers detected HPV DNA in 13.0% of OSCC cases and this rate was raise to 17.4% with the use of PGMY/GP+ nested PCR. According to our results the PGMY/GP+ nested PCR is the most appropriate primer set for the detection of HPV DNA using FFPE samples from OSCC.
authors.affiliation: Postgraduate Program in Pharmacy, Federal University of Santa Catarina, Florianópolis, Brazil. Electronic address: sibele@miyata.com.br.
authors.forename: Sibele Morais Miyata
authors.initials: SM
authors.lastname: Erhart
authors.affiliation: Department of Pathology, Federal University of Santa Catarina, Florianópolis, Brazil. Electronic address: riet.elena@gmail.com.
authors.forename: Elena Riet Correa
authors.initials: ER
authors.lastname: Rivero
authors.affiliation: Department of Clinical Analysis, Federal University of Santa Catarina, Florianópolis, Brazil. Electronic address: m.l.bazzo@ufsc.br.
authors.forename: Maria Luiza
authors.initials: ML
authors.lastname: Bazzo
authors.affiliation: Department of Clinical Analysis, Federal University of Santa Catarina, Florianópolis, Brazil. Electronic address: alexandre.onofre@gmail.com.
authors.forename: Alexandre Sherlley Casimiro
authors.initials: AS
authors.lastname: Onofre
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000175:diagnosis / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics; D006801:Humans; D009062:Mouth Neoplasms / Q000175:diagnosis / Q000235:genetics*; D027383:Papillomaviridae / Q000235:genetics*; D030361:Papillomavirus Infections / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D017931:DNA Primers; D004279:DNA, Viral
keywords: Carcinoma, squamous cell; Oncogenic viruses; Papillomavirus, human; Polymerase chain reaction