Capsid Integrity qPCR-An Azo-Dye Based and Culture-Independent Approach to Estimate Adenovirus Infectivity after Disinfection and in the Aquatic Environment

Mats Leifels; David Shoults; Alyssa Wiedemeyer; Nicholas J. Ashbolt; Emanuele Sozzi; Angela Hagemeier; Lars Jurzik

doi:10.3390/w11061196

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Capsid Integrity qPCR-An Azo-Dye Based and Culture-Independent Approach to Estimate Adenovirus Infectivity after Disinfection and in the Aquatic Environment

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Capsid Integrity qPCR-An Azo-Dye Based and Culture-Independent Approach to Estimate Adenovirus Infectivity after Disinfection and in the Aquatic Environment

Mats Leifels, David Shoults, Alyssa Wiedemeyer, Nicholas J. Ashbolt, Emanuele Sozzi, Angela Hagemeier and Lars Jurzik

Water (Basel), Vol.11(6), 1196

08/06/2019

DOI: https://doi.org/10.3390/w11061196

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Abstract

Environmental Sciences

Environmental Sciences & Ecology

Life Sciences & Biomedicine

Physical Sciences

Science & Technology

Water Resources

capsid integrity qPCR

human adenovirus

surface water

viral infectivity

water quality indicator

Recreational, reclaimed and drinking source waters worldwide are under increasing anthropogenic pressure, and often contain waterborne enteric bacterial, protozoan, and viral pathogens originating from non-point source fecal contamination. Recently, the capsid integrity (ci)-qPCR, utilizing the azo-dyes propidium monoazide (PMA) or ethidium monoazide (EMA), has been shown to reduce false-positive signals under laboratory conditions as well as in food safety applications, thus improving the qPCR estimation of virions of public health significance. The compatibility of two widely used human adenovirus (HAdV) qPCR protocols was evaluated with the addition of a PMA/EMA pretreatment using a range of spiked and environmental samples. Stock suspensions of HAdV were inactivated using heat, UV, and chlorine before being quantified by cell culture, qPCR, and ci-qPCR. Apparent inactivation of virions was detected for heat and chlorine treated HAdV while there was no significant difference between ci-qPCR and qPCR protocols after disinfection by UV. In a follow-up comparative analysis under more complex matrix conditions, 51 surface and 24 wastewater samples pre/post UV treatment were assessed for enteric waterborne HAdV to evaluate the ability of ci-qPCR to reduce the number of false-positive results when compared to conventional qPCR and cell culture. Azo-dye pretreatment of non-UV inactivated samples was shown to improve the ability of molecular HAdV quantification by reducing signals from virions with an accessible genome, thereby increasing the relevance of qPCR results for public health purposes, particularly suited to resource-limited low and middle-income settings.

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Title: Capsid Integrity qPCR-An Azo-Dye Based and Culture-Independent Approach to Estimate Adenovirus Infectivity after Disinfection and in the Aquatic Environment
Creators: Mats Leifels - Nanyang Technological University
David Shoults - University of Alberta
Alyssa Wiedemeyer - University of Alberta
Nicholas J. Ashbolt - University of Alberta
Emanuele Sozzi - University of North Carolina at Chapel Hill
Angela Hagemeier - Ruhr University Bochum
Lars Jurzik - Fed State Agcy Consumer & Hlth Protect Rhineland, D-56068 Koblenz, Germany
Publication Details: Water (Basel), Vol.11(6), 1196
Publisher: Mdpi
Number of pages: 15
Grant note: Ministry of Innovation, Science and Research North Rhine-Westphalia, Germany funding measure-Risk Management of Emerging Compounds and Pathogens in the Water Cycle (RiSKWa) 02WRS1279K / German Federal Ministry of Education and Research (BMBF); Federal Ministry of Education & Research (BMBF)
Identifiers: 991013098097402368
Copyright: © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Academic Unit: Faculty of Science and Engineering
Language: English
Resource Type: Journal article

Capsid Integrity qPCR-An Azo-Dye Based and Culture-Independent Approach to Estimate Adenovirus Infectivity after Disinfection and in the Aquatic Environment

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