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Transbound Emerg Dis ; 64(6): 1929-1934, 2017 Dec.
Article in English | MEDLINE | ID: mdl-27878975

ABSTRACT

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 TCID50 /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.


Subject(s)
Cattle Diseases/diagnosis , Coinfection/veterinary , Multiplex Polymerase Chain Reaction/veterinary , Oligonucleotide Array Sequence Analysis/veterinary , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Virus Diseases/veterinary , Animals , Cattle , Cattle Diseases/virology , Coinfection/diagnosis , Coinfection/virology , Multiplex Polymerase Chain Reaction/methods , Oligonucleotide Array Sequence Analysis/methods , Reverse Transcriptase Polymerase Chain Reaction/methods , Sensitivity and Specificity , Virus Diseases/diagnosis , Virus Diseases/virology , Viruses/isolation & purification
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