title: Immunomagnetic separation combined with RT-qPCR for determining the efficacy of disinfectants against human noroviruses.
abstract: Little is known about the effectiveness of disinfectants against human noroviruses (NoV) partially because human NoV cannot be routinely cultured in laboratory. The objective of this study was to develop a NoV monoclonal antibody-conjugated immunomagnetic separation (IMS) procedure combined with real-time reverse transcription polymerase chain reaction (RT-qPCR) assays to study the in vitro efficacy of disinfectants against human NoV. Monoclonal antibodies against Norwalk virus (NV, GI.1) and NoV GII.4 were produced using unique NoV capsid proteins, and the antibodies were conjugated to magnetic Dynalbeads. The immunomagnetic beads were used to simultaneously capture intact NoV in samples and effectively remove PCR inhibitors. We examined the efficacy of ethanol, sodium hypochlorite, nine commercially available disinfectants, and one prototype disinfectant using the IMS/RT-qPCR. The sensitivity of this procedure was approximately 100 virus particles for both the NV and GII.4 viruses. The average log reductions in in vitro activities varied between disinfectants. The prototype disinfectant produced an average 3.19-log reduction in NV and a 1.38-log reduction in GII.4. The prototype disinfectant is promising of inactivating NoV. This method can be used to evaluate in vitro activity of disinfectants against human NoV. The IMS/RT-qPCR method is promising as an effective method to remove PCR inhibitors in disinfectants and enable the evaluation of the efficacy of disinfectants.
authors.affiliation: Rollins School of Public Health, Emory University, Atlanta, GA 30322, USA. Electronic address: pliu0628@gmail.com.
authors.forename: Pengbo
authors.initials: P
authors.lastname: Liu
authors.affiliation: Kim Laboratories Inc., Rantoul, IL 61866, USA.
authors.forename: Myung
authors.initials: M
authors.lastname: Kim
authors.affiliation: Kim Laboratories Inc., Rantoul, IL 61866, USA.
authors.forename: David
authors.initials: D
authors.lastname: Schlesinger
authors.affiliation: Kim Laboratories Inc., Rantoul, IL 61866, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Kranz
authors.affiliation: Department of Food Science and Technology, Chung-Ang University, Ansung City, South Korea.
authors.forename: Sangdo
authors.initials: S
authors.lastname: Ha
authors.affiliation: Department of Food Science and Technology, Chung-Ang University, Ansung City, South Korea.
authors.forename: Jeehyoung
authors.initials: J
authors.lastname: Ha
authors.affiliation: Department of Microbiology and College of Medicine, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
authors.forename: James
authors.initials: J
authors.lastname: Slauch
authors.affiliation: Department of Food Science and Technology, Chung-Ang University, Ansung City, South Korea.
authors.forename: Seungbum
authors.initials: S
authors.lastname: Baek
authors.affiliation: Rollins School of Public Health, Emory University, Atlanta, GA 30322, USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Moe
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000302:isolation & purification; D036022:Capsid Proteins / Q000276:immunology; D004202:Disinfectants / Q000494:pharmacology*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D051379:Mice; D029322:Norovirus / Q000187:drug effects* / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D019562:Viral Load; D014773:Virology / Q000379:methods*; D038301:Virus Inactivation
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins; D004202:Disinfectants
keywords: Disinfectant; Immunomagnetic separation; Norovirus; PCR inhibition; Real-time RT-PCR