DNA Heat Treatment for Improving qPCR Analysis of Human Adenovirus in Wastewater

Food Environ Virol. 2017 Sep;9(3):354-357. doi: 10.1007/s12560-017-9294-4. Epub 2017 Apr 10.

Abstract

PCR inhibitory substances in complex sample matrices can cause false negatives or under-estimation of target concentration. This study assessed DNA heat treatment for reducing inhibition during qPCR analysis of human adenovirus (HAdV) in wastewater samples. Inhibition was reduced by heat treating DNA, where mean HAdV concentration was increased by 0.71 log10 GC/L (and up to 3.04 log10 GC/L in one case), and replicate variability and false negatives were reduced. DNA heat treatment should be further investigated for improving reliability of HAdV concentration estimates in water, which can support more accurate assessment of health risks associated with viral pathogen exposure.

Keywords: Adenovirus; DNA heat treatment; Health risk; Inhibitors; QPCR; Wastewater.

Publication types

  • Evaluation Study

MeSH terms

  • Adenoviruses, Human / classification
  • Adenoviruses, Human / genetics
  • Adenoviruses, Human / isolation & purification*
  • Analytic Sample Preparation Methods / methods*
  • DNA, Viral / chemistry
  • DNA, Viral / genetics*
  • Hot Temperature
  • Humans
  • Real-Time Polymerase Chain Reaction
  • Wastewater / virology*

Substances

  • DNA, Viral
  • Waste Water