Human Norovirus-Induced Gene Expression Biomarkers in Zebrafish.

ABSTRACT: The challenges associated with development of an animal model system to replicate human norovirus (HuNoV) has hampered the study of the pathogenesis and therapeutic interventions for this virus. In this study, we replicated HuNoV GII.4 and evaluated virus gene expression in infected zebrafish. Three doses of inoculation resulted in successful virus replication. Genes for transmembrane transporters (tfa, cftr, slc26a3, and slc26a6), a heat shock chaperone (hspa8), and immune response cytokines (ifng1 and il1b) were highly expressed in HuNoV-infected zebrafish; however, expression levels of genes were reduced in zebrafish infected with thermally inactivated HuNoV. These results confirm HuNoV replication in juvenile zebrafish and will facilitate the investigation of biomarker gene expression during HuNoV infection.

Performance of concanavalin A-immobilized on polyacrylate beads for the detection of human norovirus and hepatitis A virus in fecal specimens.

Quantitative reverse transcription PCR (qRT-PCR) is a sensitive method for the detection of foodborne viruses in fecal samples. However, the performance of qRT-PCR depends on the efficiency of virus concentration methods. In this study, the effect of Concanavalin A (Con A)-immobilized on polyacrylate beads (Con A-PAB) on the qRT-PCR performance, in terms of sensitivity and specificity to detect foodborne viruses in human fecal specimens was compared with commercial viral RNA extraction kit (VRNA). The detection of foodborne viruses by qRT-PCR was validated by viral genome sequencing. Both Con A-PAB and VRNA methods were equally sensitive and specific for detecting hepatitis A virus in fecal specimens. Even though both methods showed high specificity (100% vs. 100%) for detecting human norovirus (HuNoV), Con A-PAB method exhibited higher sensitivity (100% vs. 42.9%) and accuracy (100% vs. 73.3%) compared to VRNA method. In conclusion, the application of Con A-PAB would improve the performance of qRT-PCR for the detection of HuNoV in fecal samples.