Concentration of hepatitis A virus in milk using protamine-coated iron oxide (Fe3O4) magnetic nanoparticles

Ruiqin Wu; Xiaohui Xing; Milena Corredig; Baozhong Meng; Mansel W Griffiths

doi:10.1016/j.fm.2019.05.020

Concentration of hepatitis A virus in milk using protamine-coated iron oxide (Fe₃O₄) magnetic nanoparticles

Food Microbiol. 2019 Dec:84:103236. doi: 10.1016/j.fm.2019.05.020. Epub 2019 May 30.

Authors

Ruiqin Wu¹, Xiaohui Xing², Milena Corredig², Baozhong Meng³, Mansel W Griffiths⁴

Affiliations

¹ Department of Food Science, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada; Canadian Research Institute for Food Safety, 43 McGilvray Street, Guelph, ON, N1G 2W1, Canada.
² Department of Food Science, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada.
³ Department of Molecular and Cellular Biology, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada.
⁴ Department of Food Science, University of Guelph, 50 Stone Road East, Guelph, ON, N1G 2W1, Canada; Canadian Research Institute for Food Safety, 43 McGilvray Street, Guelph, ON, N1G 2W1, Canada. Electronic address: mgriffit@uoguelph.ca.

PMID: 31421754
DOI: 10.1016/j.fm.2019.05.020

Abstract

Hepatitis A virus (HAV) continues to be the leading cause of viral hepatitis. HAV outbreaks have been linked to the consumption of milk, but methods for HAV detection in milk are very limited. We developed a method to concentrate HAV in milk using protamine-coated iron oxide (Fe₃O₄) magnetic nanoparticles (PMNPs). In this study, protamine was covalently coated on the surface of the MNPs (20-30 nm) by a three-step chemical reaction. The successful linkage of protamine to the MNPs was confirmed by Fourier transform infrared spectroscopy (FTIR), zeta potential, and transmission electron microscopy (TEM). When used for concentrating HAV from 40 mL of milk, 50 μL of PMNPs were added to the sample and mixed for 20 min by gentle rotation, followed by a magnet capture for 30 min. The captured PMNPs were washed with glycine buffer (0.05 M glycine, 0.14 M NaCl, 0.2% (v/v) Tween 20, pH 9.0) and HAV RNA was extracted using the QIAamp MinElute Virus Spin Kit and quantified by real-time RT-PCR. The method showed a detection limit of 8.3 × 10⁰ PFU of HAV in milk. The whole concentration procedure could be completed in approximately 50 min. The developed method was simple, inexpensive, and easy-to-perform.

Keywords: Concentration; Hepatitis A virus; Iron oxide magnetic nanoparticles; Milk; Protamine.

MeSH terms

Animals
Ferric Compounds / chemistry*
Food Microbiology / methods*
Hepatitis A virus / isolation & purification*
Limit of Detection
Magnetite Nanoparticles
Milk / virology*
Protamines / chemistry*
RNA, Viral

Substances

Ferric Compounds
Magnetite Nanoparticles
Protamines
RNA, Viral
ferric oxide