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Multiplex RT ‐ PCR and Automated Microarray for Detection of Eight Bovine Viruses
Author(s) -
Lung O.,
FurukawaStoffer T.,
Burton Hughes K.,
Pasick J.,
King D. P.,
Hodko D.
Publication year - 2017
Publication title -
transboundary and emerging diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.392
H-Index - 63
eISSN - 1865-1682
pISSN - 1865-1674
DOI - 10.1111/tbed.12591
Subject(s) - agency (philosophy) , lung disease , food inspection , food safety , political science , library science , geography , medicine , pathology , lung , sociology , social science , computer science
Summary Microarrays can be a useful tool for pathogen detection as it allow for simultaneous interrogation of the presence of a large number of genetic sequences in a sample. However, conventional microarrays require extensive manual handling and multiple pieces of equipment for printing probes, hybridization, washing and signal detection. In this study, a reverse transcription ( RT )– PCR with an accompanying novel automated microarray for simultaneous detection of eight viruses that affect cattle [vesicular stomatitis virus ( VSV ), bovine viral diarrhoea virus type 1 and type 2, bovine herpesvirus 1, bluetongue virus, malignant catarrhal fever virus, rinderpest virus ( RPV ) and parapox viruses] is described. The assay accurately identified a panel of 37 strains of the target viruses and identified a mixed infection. No non‐specific reactions were observed with a panel of 23 non‐target viruses associated with livestock. Vesicular stomatitis virus was detected as early as 2 days post‐inoculation in oral swabs from experimentally infected animals. The limit of detection of the microarray assay was as low as 1 TCID 50 /ml for RPV . The novel microarray platform automates the entire post‐ PCR steps of the assay and integrates electrophoretic‐driven capture probe printing in a single user‐friendly instrument that allows array layout and assay configuration to be user‐customized on‐site.