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United States Department of Agriculture

Agricultural Research Service

Research Project: Molecular Biology of Human Pathogens Associated with Food

Location: Produce Safety and Microbiology Research

Title: Microarrays for foodborne virus detection and typing(Abstract)

Author
item Quiñones, Beatriz

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: September 25, 2012
Publication Date: N/A

Technical Abstract: Human noroviruses (HuNoV) cause up to 21 million cases of foodborne disease in the United States annually and are considered to be the most common cause of acute gastroenteritis in industrialized countries. To implement the use of simpler, rapid and cost-effective method for the simultaneous detection and genotyping of human norovirus (HuNoV), we employed the ampliPHOX colorimetric technology in conjunction with lower density microarray applications. The ampliPHOX colorimetric detection technology, based on light-initiated signal amplification through polymerization, is an inexpensive alternative to fluorescence detection and uses reagents and instrumentation that are suitable for routine pathogen surveillance. A low-density DNA microarray was designed to consist of a control biotinylated oligonucleotide and unique oligonucleotides probes(25-mer and 35-mer). The design of the unique probes was based on HuNoV genome sequences provided by the Calicivirus Laboratory with the Centers for Disease Control and Prevention. Virus-specific probes targeted regions of the HuNoV genome corresponding to the 3’-end of ORF1 (region C) and the 5’-end of ORF2(region B)of genetically-representative HuNoV GI and GII types. The microarray was designed to produce distinctly different patterns, specific for each GI and GII strain. The microarray hybridization of amplification products corresponding to regions B and C was performed according to procedures that have been optimized for detection of influenza viruses or bacterial pathogens on low density microarrays. The specific colorimetric detection of positive signals on the microarray was determined with the ampliPHOX system. Analysis of the positive signals that were detected on the microarray indicated that the use of photopolymerization(ampliPHOX colorimetric method)with low density microarrays enabled the detection of NoV genogroups and accurately genotyped genetic types belonging to a particular norovirus genogroup.

Last Modified: 8/27/2014
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