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

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.

SUBMITTER: Rames E 

PROVIDER: S-EPMC5548852 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature

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

Rames Emily E   Roiko Anne A   Stratton Helen H   Macdonald Joanne J  

Food and environmental virology 20170410 3

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 log<sub>10</sub> GC/L (and up to 3.04 log<sub>10</sub> GC/L in one case), and replicate variability and false negatives were reduced. DNA heat tr  ...[more]