title: Evaluation of commercial kits for the extraction and purification of viral nucleic acids from environmental and fecal samples.
abstract: The extraction and purification of nucleic acids is a critical step in the molecular detection of enteric viruses from environmental or fecal samples. In the present study, the performance of three commercially available kits was assessed: the MO BIO PowerViral Environmental DNA/RNA Isolation kit, the Qiagen QIAamp Viral RNA Mini kit, and the Zymo ZR Virus DNA/RNA Extraction kit. Viral particles of adenovirus 2 (AdV), murine norovirus (MNV), and poliovirus type 1 (PV1) were spiked in molecular grade water and three different types of sample matrices (i.e., biosolids, feces, and surface water concentrates), extracted with the kits, and the yields of the nucleic acids were determined by quantitative PCR (qPCR). The MO BIO kit performed the best with the biosolids, which were considered to contain the highest level of inhibitors and provided the most consistent detection of spiked virus from all of the samples. A qPCR inhibition test using an internal control plasmid DNA and a nucleic acid purity test using an absorbance at 230 nm for the nucleic acid extracts demonstrated that the MO BIO kit was able to remove qPCR inhibitors more effectively than the Qiagen and Zymo kits. These results suggest that the MO BIO kit is appropriate for the extraction and purification of viral nucleic acids from environmental and clinical samples that contain high levels of inhibitors.
authors.affiliation: Department of Soil, Water and Environmental Science, The University of Arizona, Tucson, AZ 85721, USA.
authors.forename: Brandon C
authors.initials: BC
authors.lastname: Iker
authors.forename: Kelly R
authors.initials: KR
authors.lastname: Bright
authors.forename: Ian L
authors.initials: IL
authors.lastname: Pepper
authors.forename: Charles P
authors.initials: CP
authors.lastname: Gerba
authors.forename: Masaaki
authors.initials: M
authors.lastname: Kitajima
mesh_terms: D000818:Animals; D004267:DNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D004783:Environmental Microbiology; D005243:Feces / Q000821:virology*; D006801:Humans; D009696:Nucleic Acids / Q000235:genetics / Q000302:isolation & purification*; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction; D014773:Virology / Q000379:methods*
chemical_list: D009696:Nucleic Acids