Main content

Development of Novel Methods for the Concentration and Detection of Foodborne Hepatitis A virus

Show simple item record

dc.contributor.advisor Griffiths, Mansel
dc.contributor.author Wu, Ruiqin
dc.date.accessioned 2014-11-18T21:17:36Z
dc.date.available 2015-10-31T05:00:12Z
dc.date.copyright 2014-10
dc.date.created 2014-10-31
dc.date.issued 2014-11-18
dc.identifier.uri http://hdl.handle.net/10214/8542
dc.description.abstract The low amounts of viral particles in contaminated foods make it challenging to develop methods for their detection. Rapid and easy-to-use methods for separating and concentrating viruses from contaminated foods are needed to enhance the efficiency of virus detection. In the present study, iron oxide (Fe3O4) magnetic nanoparticles (MNPs) coated with -NH2 or protamine (designated NMNPs and PMNPs, respectively) were used to concentrate hepatitis A virus (HAV) from different food matrices. Successful coating of -NH2 or protamine was confirmed using Fourier transform infrared spectroscopy (FTIR), zeta potential test, and transmission electron microscopy (TEM). Glycine buffer (0.05 M glycine, 0.14 M NaCl, 0.2% (v/v) Tween 20, pH 9.0) was found to be efficient in eluting HAV from artificially contaminated green onion, strawberry, and mussel. When used for concentrating HAV from the food eluates or milk, PMNPs showed higher sensitivity and repeatability than NMNPs. The detection limit of HAV by real-time RT-PCR was 8.3 plaque-forming unit (PFU)/15 g, 83 PFU/50 g, 8.3 PFU/5 g, and 8.3 PFU/40 mL for green onion, strawberry, mussel, and milk, respectively. Polyethylene glycol (PEG) dialysis method demonstrated similar analytical sensitivity to the PMNP method in concentrating HAV from green onion, mussel, and milk. A bioluminescent real-time reverse transcriptase loop-mediated isothermal amplification (RT-LAMP-BART) technology was employed in the current study to detect HAV concentrated from green onion, strawberry, mussel, and milk by PMNPs. The analytical sensitivity of cDNA-LAMP-BART assay was 8.3 PFU/15 g, 83 PFU/50 g, 8.3 PFU/5 g, and 8.3 PFU/40 mL for green onion, strawberry, mussel, and milk, respectively. These results were comparable with those from real-time RT-PCR. Qβ replicase reaction assay was also investigated in the present study for HAV detection. It was found that the method was not applicable in detecting foodborne virus due to the strong background signal in non-template reactions, which gave false positive results. en_US
dc.language.iso en en_US
dc.subject Hepatitis A Virus en_US
dc.subject Concentration en_US
dc.subject Detection en_US
dc.subject Magnetic Nanoparticles en_US
dc.subject RT-LAMP-BART en_US
dc.title Development of Novel Methods for the Concentration and Detection of Foodborne Hepatitis A virus en_US
dc.type Thesis en_US
dc.degree.programme Food Science en_US
dc.degree.name Doctor of Philosophy en_US
dc.degree.department Department of Food Science en_US
dc.rights.license All items in the Atrium are protected by copyright with all rights reserved unless otherwise indicated.


Files in this item

Files Size Format View Description
Wu_Ruiqin_2014_10_PhD.pdf 29.36Mb PDF View/Open Thesis

This item appears in the following Collection(s)

Show simple item record