Electronic microarray assays for avian influenza and Newcastle disease virus.
Microarrays are suitable for multiplexed detection and typing of pathogens. Avian influenza virus (AIV) is currently classified into 16 H (hemagglutinin) and 9 N (neuraminidase) subtypes, whereas Newcastle disease virus (NDV) strains differ in virulence and are broadly classified into high and low pathogenicity types. In this study, three assays for detection and typing of poultry viruses were developed on an automated microarray platform: a multiplex assay for simultaneous detection of AIV and detection and pathotyping of NDV, and two separate assays for differentiating all AIV H and N subtypes. The AIV-NDV multiplex assay detected all strains in a 63 virus panel, and accurately typed all high pathogenicity NDV strains tested. A limit of detection of 10(1)-10(3) TCID(50)/mL and 200-400 EID(50)/mL was obtained for NDV and AIV, respectively. The AIV typing assays accurately typed all 41 AIV strains and a limit of detection of 4-200 EID(50)/mL was obtained. Assay validation showed that the microarray assays were generally comparable to real-time RT-PCR. However, the AIV typing microarray assays detected more positive clinical samples than the AIV matrix real-time RT-PCR, and also provided information regarding the subtype. The AIV-NDV multiplex and AIV H typing microarray assays detected mixed infections and could be useful for detection and typing of AIV and NDV.
Canadian Food Inspection Agency, National Centres for Animal Disease, Lethbridge Laboratory, Township Road 9-1, P.O. Box 640, Lethbridge, Alberta T1J 3Z4, Canada. Oliver.Lung@inspection.gc.ca
Oliver
O
Lung
Anne
A
Beeston
Samuel
S
Ohene-Adjei
John
J
Pasick
Dalibor
D
Hodko
Kimberley Burton
KB
Hughes
Tara
T
Furukawa-Stoffer
Mathew
M
Fisher
Dirk
D
Deregt
D000818:Animals; D002645:Chickens / Q000821:virology; D005814:Genes, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000032:analysis / Q000235:genetics; D009980:Influenza A virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology; D009439:Neuraminidase / Q000032:analysis / Q000235:genetics; D009521:Newcastle Disease / Q000175:diagnosis* / Q000821:virology; D009522:Newcastle disease virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods / Q000662:veterinary*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D012680:Sensitivity and Specificity; D014422:Turkeys / Q000821:virology; D014760:Viral Fusion Proteins / Q000032:analysis / Q000235:genetics
D019267:Hemagglutinin Glycoproteins, Influenza Virus; D014760:Viral Fusion Proteins; D009439:Neuraminidase