Evaluation of three different concentration and extraction methods for recovery efficiency of human adenovirus and human rotavirus virus A

Islam Nour; Atif Hanif; Fahd Alanazi; Adel M Zakri; Ibrahim Al-Ashkar; Abdulkarim Alhetheel; Saleh Eifan

doi:10.1016/j.jviromet.2021.114212

Evaluation of three different concentration and extraction methods for recovery efficiency of human adenovirus and human rotavirus virus A

J Virol Methods. 2021 Sep:295:114212. doi: 10.1016/j.jviromet.2021.114212. Epub 2021 Jun 11.

Authors

Islam Nour¹, Atif Hanif², Fahd Alanazi¹, Adel M Zakri³, Ibrahim Al-Ashkar³, Abdulkarim Alhetheel⁴, Saleh Eifan¹

Affiliations

¹ Botany and Microbiology Department, College of Science, King Saud University, Saudi Arabia.
² Botany and Microbiology Department, College of Science, King Saud University, Saudi Arabia. Electronic address: ahchaudhry@ksu.edu.sa.
³ Biotechnology Laboratory, Plant Production Department, Faculty of Agriculture and Food Sciences, King Saud University, Saudi Arabia.
⁴ Department of Pathology and Laboratory Medicine, College of Medicine, King Saud University, Saudi Arabia.

PMID: 34126107
DOI: 10.1016/j.jviromet.2021.114212

Abstract

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.

Keywords: Adenovirus; Concentration; Extraction; Rotavirus A; Treated wastewater; Virus recovery.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenoviruses, Human* / genetics
Enterovirus*
Humans
Real-Time Polymerase Chain Reaction
Rotavirus* / genetics
Water Purification*