title: Automated high throughput DNA isolation for detection of human papillomavirus in oral rinse samples.
abstract: BACKGROUND
  Oral HPV infection elevates risk of oropharyngeal cancer, but its natural history is unknown. Natural history studies necessitate validation of an automated, high-throughput method for HPV genomic DNA detection in oral rinse samples (ORS).
  
  
  OBJECTIVE
  To compare agreement of oral HPV detection in ORS processed by a magnetic-bead based automated platform to a previous gold-standard, manual protein-precipitation method. Agreement was compared to that of repeat sampling and repeat HPV testing.
  
  
  METHODS
  HIV-infected individuals (n=100) provided two ORS collected 15 min apart. DNA was isolated from equal aliquots by either a protein-precipitation based (Puregene, Qiagen) or magnetic bead-based (QIAsymphony™ SP, Qiagen) method. HPV DNA was detected and type-specified by consensus primer PCR and reverse line blot hybridization. The kappa statistic was used to assess overall agreement (OA) and agreement on a positive test (Ps+).
  
  
  RESULTS
  The DNA purification methods had very high agreement for categorizing an individual as HPV infected (OA=0.95; Ps+=0.94) as well as for detection of HPV type-specific infection (OA=0.99; Ps+=0.88) in ORS. Agreement for detection of HPV type-specific infection was greater than that observed with repeat oral rinse sampling (OA=0.99, Ps+=0.76) but comparable to inter-assay agreement (OA=1.00, Ps+=0.90).
  
  
  CONCLUSIONS
  HPV detection in ORS processed with a magnetic-bead based automated platform will facilitate large natural history studies of oral HPV infection necessary to evaluate the potential use of oral HPV detection in oral cancer screening.
authors.affiliation: Viral Oncology, Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Tatevik R
authors.initials: TR
authors.lastname: Broutian
authors.forename: Xin
authors.initials: X
authors.lastname: He
authors.forename: Maura L
authors.initials: ML
authors.lastname: Gillison
mesh_terms: D015331:Cohort Studies; D004279:DNA, Viral / Q000032:analysis / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009055:Mouth / Q000821:virology*; D009059:Mouth Diseases / Q000821:virology*; D009067:Mouthwashes; D027383:Papillomaviridae / Q000235:genetics* / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012307:Risk Factors
chemical_list: D004279:DNA, Viral; D009067:Mouthwashes