Development of Electronic Microarray Assays for the Detection of High Consequence Swine Viruses

Abstract

This thesis describes the development and optimization of a seven-plex reverse transcription polymerase chain reaction (RT-PCR) and corresponding user-friendly electronic microarray for the detection of seven swine viruses: foot-and-mouth disease, swine vesicular disease, classical swine fever, vesicular exanthema of swine, African swine fever, porcine circovirus type 2 and porcine reproductive and respiratory syndrome. A panel of 58 strains of the viruses were successfully amplified and detected specifically on the NanoChip 400 microarray system while having no detection of 22 non-specific clinical material and non-target viruses and bacteria. Target viruses were also detected from clinical and biological materials spiked with viruses as early as 1 days post-infection. Detection limits ranged from 10 to 1000 copies for the targets. The assay was successfully transferred to the prototype Nexogen MDx portable and integrated system, where the fully automated system, with no manual handling from sample to detection, detected five of the seven viruses.