title: Capture and concentration of viral and bacterial foodborne pathogens using apolipoprotein H.
abstract: The need for improved pathogen separation and concentration methods to reduce time-to-detection for foodborne pathogens is well recognized. Apolipoprotein H (ApoH) is an acute phase human plasma protein that has been previously shown to interact with viruses, lipopolysaccharides (LPS) and bacterial proteins. The purpose of this study was to determine if ApoH was capable of binding and efficiently capturing two representative human norovirus strains (GI.1 and GII.4), a cultivable surrogate, and four bacterial pathogens (Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica serovar Enteritidis, and Staphylococcus aureus). Experiments were carried out using an ApoH-conjugated magnetic bead-based capture followed by pathogen detection using nucleic acid amplification. For all three viruses studied, >10% capture efficiency (<1 Log10 loss in RT-qPCR amplifiable units) was observed. The same capture efficiencies were observed for the bacterial pathogens tested, with the exception of E. coli O157:H7 (approximately 1% capture efficiency, or 2 Log10 loss in CFU equivalents). The efficiency of the capture steps did not vary as a consequence of input target concentration or in the presence of an abundance of background microflora. A complementary plate-based capture assay showed that ApoH bound to a variety of human norovirus virus-like particles. ApoH has the potential to be a broadly reactive ligand for separating and concentrating representative foodborne pathogens, both bacteria and viruses.
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC 27695, USA. Electronic address: eaalmand@ncsu.edu.
authors.forename: Erin A
authors.initials: EA
authors.lastname: Almand
authors.affiliation: Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC 27695, USA.
authors.forename: Rebecca M
authors.initials: RM
authors.lastname: Goulter
authors.affiliation: Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC 27695, USA; Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC 27695, USA. Electronic address: lajaykus@ncsu.edu.
authors.forename: Lee-Ann
authors.initials: LA
authors.lastname: Jaykus
mesh_terms: D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000302:isolation & purification*; D019453:Escherichia coli O157 / Q000302:isolation & purification; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D005517:Foodborne Diseases / Q000175:diagnosis* / Q000382:microbiology / Q000821:virology; D008089:Listeria monocytogenes / Q000302:isolation & purification; D009663:Norwalk virus / Q000302:isolation & purification; D012367:RNA, Viral / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction; D012477:Salmonella enteritidis / Q000302:isolation & purification; D017423:Sequence Analysis, RNA; D013211:Staphylococcus aureus / Q000302:isolation & purification; D053482:beta 2-Glycoprotein I / Q000737:chemistry*
chemical_list: D004269:DNA, Bacterial; D012367:RNA, Viral; D053482:beta 2-Glycoprotein I
keywords: ApoH; Bacteria; Concentration; Detection; Norovirus