Microarray-based detection and typing of foot-and-mouth disease virus.
Foot-and-mouth disease virus (FMDV) is the most economically important veterinary pathogen because of its highly infectious nature and the devastating effects the virus has on the livestock industry. Rapid diagnostic methods are needed for detection and typing of FMDV serotypes and differentiation from other viruses causing vesicular diseases. We developed a microarray-based test that uses a FMD DNA chip containing 155 oligonucleotide probes, 35-45 base pair (bp) long, virus-common and serotype-specific, designed from the VP3-VP1-2A region of the genome. A set of two forward primers and one reverse primer were also designed to allow amplification of approximately 1100 bp of target sequences from this region. The amplified target was labelled with Alexa-Fluor 546 dye and applied to the FMD DNA chip. A total of 23 different FMDV strains representing all seven serotypes were detected and typed by the FMD DNA chip. Microarray technology offers a unique capability to identify multiple pathogens in a single chip.
Baxi
Mohit K
MK
Virology Section, Lethbridge Laboratory (Animal Diseases Research Institute), Canadian Food Inspection Agency, Lethbridge, Alberta, Canada. baxim@inspection.gc.ca
Baxi
Shailja
S
Clavijo
Alfonso
A
Burton
Kimberly M
KM
Deregt
Dirk
D
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D005536:Foot-and-Mouth Disease / Q000175:diagnosis / Q000821:virology*; D029544:Foot-and-Mouth Disease Virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods / Q000662:veterinary; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012756:Sheep
D012367:RNA, Viral