Capsid-Damaging Effects of UV Irradiation as Measured by Quantitative PCR Coupled with Ethidium Monoazide Treatment

Food Environ Virol. 2014 Dec;6(4):269-75. doi: 10.1007/s12560-014-9162-4. Epub 2014 Aug 9.

Abstract

The damage to a viral capsid after low-pressure (LP) and medium-pressure (MP) UV irradiation was assessed, using the quantitative or quantitative reverse transcription PCR coupled with ethidium monoazide treatment (EMA-PCR). After UV irradiation, adenovirus 5 (Ad5) and poliovirus 1 (PV1) were subjected to a plaque assay, PCR, and EMA-PCR to investigate the effect of UV irradiation on viral infectivity, genome damage, and capsid damage, respectively. The effectiveness of UV wavelengths in a viral genome and capsid damage of both PV1 and Ad5 was also further investigated using a band-pass filter. It was found that an MPUV lamp was more effective than an LPUV lamp in inactivating Ad5, whereas there was no difference in the case of PV1. The results of viral reduction determined by PCR and EMA-PCR indicated that MP UV irradiation damaged Ad5 capsid. The damage to PV1 and Ad5 capsid was also not observed after LP UV irradiation. The investigation of effects of UV wavelengths suggested that UV wavelengths at 230-245 nm have greater effects on adenovirus capsid in addition to viral genome than UV wavelengths beyond 245 nm.

Keywords: Ethidium monoazide; UV irradiation; Viral capsid damage; Viral genome damage.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviruses, Human / growth & development
  • Adenoviruses, Human / metabolism
  • Adenoviruses, Human / pathogenicity
  • Adenoviruses, Human / radiation effects*
  • Affinity Labels / pharmacology*
  • Animals
  • Azides / pharmacology*
  • Capsid / metabolism
  • Capsid / radiation effects*
  • Cell Line
  • Chlorocebus aethiops
  • DNA, Viral / metabolism
  • DNA, Viral / radiation effects
  • Disinfection / methods*
  • Genome, Viral / radiation effects*
  • Humans
  • Poliovirus / growth & development
  • Poliovirus / metabolism
  • Poliovirus / pathogenicity
  • Poliovirus / radiation effects*
  • Pressure
  • RNA, Viral / metabolism
  • RNA, Viral / radiation effects
  • Radiation Tolerance
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Ultraviolet Rays
  • Viral Plaque Assay
  • Virus Inactivation / radiation effects

Substances

  • Affinity Labels
  • Azides
  • DNA, Viral
  • RNA, Viral
  • 8-azidoethidium