title: Detecting human papillomavirus in ocular surface diseases.
abstract: OBJECTIVE
  Human papillomavirus (HPV) infection has been implicated as a possible inducing factor for benign and neoplastic ocular surface diseases such as pterygia and ocular-surface squamous neoplasia (OSSN). However, the wide range in HPV prevalence previously reported for both diseases adds controversy to, and highlights the limitations of, this field. The aim of this study was to determine the prevalence of HPV in pterygia and OSSN and to devise a standardized approach for detecting viral DNA in ocular tissue samples.
  
  
  METHODS
  DNA was extracted from a variety of specimens (n = 160), including formalin-fixed paraffin-embedded tissue shavings, fresh tissue, and cultured cells. Nested PCR for HPV with consensus and subtype-specific primers was used to detect viral DNA. Confirmatory assays, including molecular sequencing, histology, and immunohistochemistry for HPV E6 protein and p16 were also performed.
  
  
  RESULTS
  HPV was not detected in pterygia or normal conjunctiva. However, 6.5% (3/46) of OSSN samples were HPV-positive by PCR, sequencing, and immunohistochemistry. Positive cases were all squamous cell carcinoma of the conjunctiva (SCCC), the most severe form of OSSN, representing 12.5% (3/24) of SCCCs in our cohort. HPV-16 was the genotype identified in each case and this correlated with the presence of koilocytes and intense immunoreactivity for p16. Our study found no association between pterygia and OSSN with other oncogenic viruses, such as EBV or CMV, as they were just as prevalent in normal conjunctiva.
  
  
  CONCLUSIONS
  The low prevalence of HPV-16 in ocular surface disease suggests infection is not a cause but a cofactor in disease development.
authors.affiliation: School of Medical Sciences, University of New South Wales, Sydney, Australia.
authors.forename: Mark
authors.initials: M
authors.lastname: Woods
authors.forename: Sharron
authors.initials: S
authors.lastname: Chow
authors.forename: Benjamin
authors.initials: B
authors.lastname: Heng
authors.forename: Wendy
authors.initials: W
authors.lastname: Glenn
authors.forename: Noel
authors.initials: N
authors.lastname: Whitaker
authors.forename: Dale
authors.initials: D
authors.lastname: Waring
authors.forename: Jenna
authors.initials: J
authors.lastname: Iwasenko
authors.forename: William
authors.initials: W
authors.lastname: Rawlinson
authors.forename: Minas T
authors.initials: MT
authors.lastname: Coroneo
authors.forename: Denis
authors.initials: D
authors.lastname: Wakefield
authors.forename: Nick
authors.initials: N
authors.lastname: Di Girolamo
mesh_terms: D000368:Aged; D002294:Carcinoma, Squamous Cell / Q000453:epidemiology / Q000473:pathology / Q000821:virology*; D002478:Cells, Cultured; D003228:Conjunctiva / Q000166:cytology / Q000821:virology; D003230:Conjunctival Neoplasms / Q000453:epidemiology / Q000473:pathology / Q000821:virology*; D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000821:virology; D015828:Eye Infections, Viral / Q000453:epidemiology / Q000473:pathology / Q000821:virology*; D005260:Female; D052162:Human papillomavirus 16 / Q000302:isolation & purification*; D052161:Human papillomavirus 18 / Q000302:isolation & purification; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008875:Middle Aged; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism; D030361:Papillomavirus Infections / Q000453:epidemiology / Q000473:pathology / Q000821:virology*; D016133:Polymerase Chain Reaction; D015995:Prevalence; D011625:Pterygium / Q000453:epidemiology / Q000473:pathology / Q000821:virology*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D004279:DNA, Viral; D004268:DNA-Binding Proteins; C068651:E6 protein, Human papillomavirus type 16; C052603:E6 protein, Human papillomavirus type 18; D009856:Oncogene Proteins, Viral; D012097:Repressor Proteins
keywords: conjunctiva; epithelium; neoplasia; ocular surface; pterygia; virus