Detection of Human Papillomavirus Infections at the Single-Cell Level

Intervirology. 2015;58(5):324-331. doi: 10.1159/000442573. Epub 2016 Jan 28.

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.

Conclusion: 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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cervix Uteri / virology
  • DNA, Viral / analysis*
  • DNA, Viral / genetics
  • Epithelial Cells / virology*
  • Female
  • Humans
  • Laser Capture Microdissection
  • Papillomaviridae / genetics
  • Papillomaviridae / isolation & purification*
  • Papillomavirus Infections / virology*
  • Real-Time Polymerase Chain Reaction
  • Single-Cell Analysis / methods*

Substances

  • DNA, Viral