title: Detection of Human Papillomavirus Infections at the Single-Cell Level.
abstract: OBJECTIVE
  To explore the possibility of single-cell analysis of human papillomavirus (HPV) infection.
  
  
  METHODS
  Two hundred and twenty cells were isolated by laser capture microdissection from formalin-fixed and paraffin-embedded cervical tissue blocks from 8 women who had HPV DNA detected in their cervical swab samples. The number of type-specific HPV copies in individual cells was measured by quantitative polymerase chain reaction with and without a prior reverse transcription. The cells were assayed and counted for more than once if the corresponding swab sample was positive for ≥2 HPV types.
  
  
  RESULTS
  Infection with HPV16, HPV39, HPV51, HPV52, HPV58, HPV59 and HPV73 was detected in 12 (5.5%) of 220, 3 (9.4%) of 32, 3 (5.8%) of 52, 11 (22.9%) of 48, 9 (18.8%) of 48, 3 (9.4%) of 32 and none of 20 cells, respectively. The numbers of HPV genome copies varied widely from cell to cell. The coexistence of multiple HPV types was detected in 6 (31.6%) of 19 positive cells from 1 of the 6 women who had 2 or 3 HPV types detected in their swab samples.
  
  
  CONCLUSIONS
  Given the heterogeneity of HPV status in individual cells, further clarification of HPV infection at the single-cell level may refine our understanding of HPV-related carcinogenesis.
authors.affiliation: Department of Pathology, School of Medicine, University of Washington, Seattle, WA.
authors.forename: Zhenping
authors.initials: Z
authors.lastname: Shen
authors.affiliation: Department of Pathology, School of Medicine, University of Washington, Seattle, WA.|Departments of Gynecology and Obstetrics, Shengjing Hospital of China Medical University, Shenyang, Liaoning, P.R.China.
authors.forename: Xia
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Pathology, School of Medicine, University of Washington, Seattle, WA.
authors.forename: Janice
authors.initials: J
authors.lastname: Morihara
authors.affiliation: Department of Pathology, School of Medicine, University of Washington, Seattle, WA.
authors.forename: Ayaka
authors.initials: A
authors.lastname: Hulbert
authors.affiliation: Department of Epidemiology, School of Public Health, University of Washington, Seattle, WA.
authors.forename: Laura A
authors.initials: LA
authors.lastname: Koutsky
authors.affiliation: Department of Pathology, School of Medicine, University of Washington, Seattle, WA.
authors.forename: Nancy B
authors.initials: NB
authors.lastname: Kiviat
authors.affiliation: Department of Pathology, School of Medicine, University of Washington, Seattle, WA.|Department of Epidemiology, School of Public Health, University of Washington, Seattle, WA.
authors.forename: Long Fu
authors.initials: LF
authors.lastname: Xi
mesh_terms: D002584:Cervix Uteri / Q000821:virology; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D004847:Epithelial Cells / Q000821:virology*; D005260:Female; D006801:Humans; D061346:Laser Capture Microdissection; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D004279:DNA, Viral
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