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Capability of FDA-EVIR Microarray for Detection of Norovirus and Hepatitis A Virus in Inoculated Tomatoes, Green Onions, and Celery

ABSTRACT: The capability of the U.S. Food and Drug Administration Enteric Viruses tiling microarray (FDA-EVIR) was assessed for rapid molecular identification of human norovirus (NoV) and hepatitis A virus (HAV) extracted from artificially inoculated fresh produce. Two published viral extraction strategies, total RNA extraction or virus particle isolation, were employed to prepare the viral targets. We also assessed the amount of viral RNA extracted from celery by three commercially-available kits and how well that RNA performed on the FDA-EVIR. Our results confirm that FDA-EVIR can correctly identify common enteric viruses isolated from fresh produce and is capable of identifying single and mixed species of viruses, as well as distinguishing among genotypes. Extending microarray methods to other food matrices should provide important support to surveillance and outbreak investigations.

ORGANISM(S): Paslahepevirus balayani Apium graveolens Coxsackievirus Sapovirus Norovirus Solanum lycopersicum Rotavirus Bacillus subtilis Hepatovirus A Allium fistulosum unidentified adenovirus Astrovirus sp.

PROVIDER: GSE140351 | GEO | 2020/04/01

REPOSITORIES: GEO

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Project description:Hepatitis A virus (HAV) is a hepatotropic human picornavirus that has been associated only with acute infection. Its pathogenesis is not well understood since there have been few recent studies in animal models using modern methodologies. We characterized HAV infections in three chimpanzees, quantifying viral RNA by qRT-PCR and examining critical aspects of the innate immune response including intrahepatic interferon-stimulated gene expression. We compared these infection profiles with similar studies of chimpanzees infected with hepatitis C virus (HCV), a hepatotropic flavivirus that frequently causes persistent infection. Surprisingly, HAV-infected animals exhibited very limited induction of type I interferon-stimulated genes in the liver compared to chimpanzees with acute resolving HCV infection, despite similar levels of viremia and 100-fold greater quantities of viral RNA in the liver. Minimal ISG15 and IFIT1 responses peaked 1-2 weeks after HAV challenge, then subsided despite continuing high hepatic viral loads. An acute inflammatory response at 3-4 weeks correlated with the appearance of virus-specific antibodies, and both apoptosis and proliferation of hepatocytes. Despite this, HAV RNA persisted in the liver for months, remaining present long after its clearance from serum and feces and revealing dramatic differences in the kinetics of clearance in the three compartments. Viral RNA was detected in the liver for significantly longer (35 to >48 weeks) than HCV RNA in animals with acute resolving HCV infection (10-20 weeks). Collectively, these findings suggest that early HAV infection is far stealthier than HCV infection and represents a distinctly different paradigm in viral-host interactions within the liver. Chimpanzee liver was biopsied during an acute HAV infection. Chimp 1 and 2 had two baseline samples. Chimp 3 used the baselines from chimps 1 and 2. Chimp 1 had 8 samples during the HAV acute infection. Chimp 2 had 9 samples during the HAV acute infection. Chimp 3 had 4 samples during the HAV acute infection.

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Capability of FDA-EVIR Microarray for Detection of Norovirus and Hepatitis A Virus in Inoculated Tomatoes, Green Onions, and Celery

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