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From Lab to Lake – Evaluation of Current Molecular Methods for the Detection of Infectious Enteric Viruses in Complex Water Matrices in an Urban Area

View Article: PubMed Central - PubMed

ABSTRACT

Quantitative PCR methods are commonly used to monitor enteric viruses in the aquatic environment because of their high sensitivity, short reaction times and relatively low operational cost. However, conclusions for public health drawn from results of such molecular techniques are limited due to their inability to determine viral infectivity. Ethidium monoazide (EMA) and propidium monoazide (PMA) are capable to penetrate the damaged or compromised capsid of the inactivated viruses and bind to the viral nucleic acids. We assessed whether dye treatment is a suitable approach to improve the ability of qPCR to distinguish between infectious and non-infectious human adenovirus, enterovirus and rotavirus A in surface water of an urban river and sewage before and after UV disinfection. Like the gold standard of cell culture assays, pretreatment EMA-/PMA-qPCR succeeded in removing false positive results which would lead to an overestimation of the viral load if only qPCR of the environmental samples was considered. A dye pretreatment could therefore provide a rapid and relatively inexpensive tool to improve the efficacy of molecular quantification methods in regards to viral infectivity.

No MeSH data available.


Boxplot comparison of the calculated concentration for rotavirus A originating from surface waters and sewage waters before and after UV treatment as well as total samples.
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pone.0167105.g004: Boxplot comparison of the calculated concentration for rotavirus A originating from surface waters and sewage waters before and after UV treatment as well as total samples.

Mentions: RT-qPCR showed a slightly higher occurrence and median genomic concentration per liter of RV compared to EV in all samples (see Fig 4). Almost 20% of the combined, surface and sewage water samples were positive using this method. Pretreatment with EMA lead to negative RV signals in the water samples regardless of the source while only small numbers of the combined (3/79), surface (2/53) and sewage water samples (1/26; only inflow) were positive for PMA treated samples.


From Lab to Lake – Evaluation of Current Molecular Methods for the Detection of Infectious Enteric Viruses in Complex Water Matrices in an Urban Area
Boxplot comparison of the calculated concentration for rotavirus A originating from surface waters and sewage waters before and after UV treatment as well as total samples.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5120873&req=5

pone.0167105.g004: Boxplot comparison of the calculated concentration for rotavirus A originating from surface waters and sewage waters before and after UV treatment as well as total samples.
Mentions: RT-qPCR showed a slightly higher occurrence and median genomic concentration per liter of RV compared to EV in all samples (see Fig 4). Almost 20% of the combined, surface and sewage water samples were positive using this method. Pretreatment with EMA lead to negative RV signals in the water samples regardless of the source while only small numbers of the combined (3/79), surface (2/53) and sewage water samples (1/26; only inflow) were positive for PMA treated samples.

View Article: PubMed Central - PubMed

ABSTRACT

Quantitative PCR methods are commonly used to monitor enteric viruses in the aquatic environment because of their high sensitivity, short reaction times and relatively low operational cost. However, conclusions for public health drawn from results of such molecular techniques are limited due to their inability to determine viral infectivity. Ethidium monoazide (EMA) and propidium monoazide (PMA) are capable to penetrate the damaged or compromised capsid of the inactivated viruses and bind to the viral nucleic acids. We assessed whether dye treatment is a suitable approach to improve the ability of qPCR to distinguish between infectious and non-infectious human adenovirus, enterovirus and rotavirus A in surface water of an urban river and sewage before and after UV disinfection. Like the gold standard of cell culture assays, pretreatment EMA-/PMA-qPCR succeeded in removing false positive results which would lead to an overestimation of the viral load if only qPCR of the environmental samples was considered. A dye pretreatment could therefore provide a rapid and relatively inexpensive tool to improve the efficacy of molecular quantification methods in regards to viral infectivity.

No MeSH data available.