Abstract
RVA: Human Rotavirus A, HAdV: Human Adenovirus, ARQ: Armored RNA® Quant, GW: Glass wool-mediated concentration method, PEG: Polyethylene glycol percipitaion-based concentration method, CMAE: Charged membrane Adsorption/elution concentration method.
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•We established a full factorial design of 9 concentration x extraction combinations.•Significant differences for the concentrations and recovery efficiencies were noted.•ARQ Process control emphasized high concentration and extraction recoveries.•Highest HAdV recovery efficiencies were obtained by PEG-POW methods.•Highest RVA recovery efficiencies were attained by CMAE-ZYMO methods.
Routine wastewater treatment plants (WWTPs) effluents monitoring is essential because of enteric viruses’ low infectious dose beyond molecular detectability. In current study methods for concentration and extraction, inter-method compatibility and recovery efficiency of spiked human adenovirus (HAdV) and human rotavirus A (RVA) were evaluated. For virus concentration, polyethylene glycol precipitation (PEG), charged membrane-based adsorption/elution (CMAE), and glass wool-based concentration (GW) methods were used. Nucleic acid was extracted by PowerViral™ Environmental RNA/DNA Isolation (POW), ZymoBIOMICS™ RNA extraction (ZYMO) and Wizard® Genomic DNA Purification (WGDP) and samples were analyzed by Real-Time PCR. CMAE method yielded significantly higher concentrations for both ARQ (Armored-RNA Quant) and RVA compared to PEG (P = 0.001 and 0.003) and GW (P < 0.0001). Highest HAdV concentration was obtained by PEG (P = 0.001 and < 0.0001) in relation to CMAE and GW. ZYMO yielded a significantly higher ARQ and RVA concentrations (P = 0.03 and 0.0057), whereas significantly higher concentration was obtained by POW for HAdV (P = 0.032). CMAE × ZYMO achieved the highest recovery efficiencies for ARQ (69.77 %) and RVA (64.25, respectively, while PEG × POW present efficiency of 9.7 % for HAdV. These findings provide guidance for understanding of method-related biases for viral recovery efficiency.