Electronic microarray assays for avian influenza and Newcastle disease virus

► 3 PCR-based microarray assays were developed for detection and typing of AIV and NDV. ► Assay 1 detects AIV/NDV and pathotypes NDV. Assays 2 and 3 identify AIV H- and N-types. ► Mixed infections were detected by the multiplexed assay and the AIV H-typing assay. Microarrays are suitable for multipl...

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Bibliographic Details
Published in:Journal of virological methods 2012-11, Vol.185 (2), p.244-253
Main Authors: Lung, Oliver, Beeston, Anne, Ohene-Adjei, Samuel, Pasick, John, Hodko, Dalibor, Hughes, Kimberley Burton, Furukawa-Stoffer, Tara, Fisher, Mathew, Deregt, Dirk
Format: Article
Language:English
Subjects:
Animals
avian influenza
Avian influenza virus
Biological and medical sciences
Chickens - virology
detection limit
Fundamental and applied biological sciences. Psychology
Genes, Viral
Hemagglutinin Glycoproteins, Influenza Virus - analysis
Hemagglutinin Glycoproteins, Influenza Virus - genetics
hemagglutinins
Influenza
Influenza A virus
Influenza A virus - classification
Influenza A virus - genetics
Influenza A virus - isolation & purification
Influenza in Birds - diagnosis
Influenza in Birds - virology
Microarray
microarray technology
Microbiology
mixed infection
Multiplex PCR
Neuraminidase - analysis
Neuraminidase - genetics
Newcastle disease
Newcastle Disease - diagnosis
Newcastle Disease - virology
Newcastle disease virus
Newcastle disease virus - classification
Newcastle disease virus - genetics
Newcastle disease virus - isolation & purification
Oligonucleotide Array Sequence Analysis - methods
Oligonucleotide Array Sequence Analysis - veterinary
pathogens
poultry
reverse transcriptase polymerase chain reaction
Reverse Transcriptase Polymerase Chain Reaction - methods
Reverse Transcriptase Polymerase Chain Reaction - veterinary
Sensitivity and Specificity
sialidase
Techniques used in virology
Turkeys - virology
Typing
Viral Fusion Proteins - analysis
Viral Fusion Proteins - genetics
Virology
virulence
viruses
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Summary:► 3 PCR-based microarray assays were developed for detection and typing of AIV and NDV. ► Assay 1 detects AIV/NDV and pathotypes NDV. Assays 2 and 3 identify AIV H- and N-types. ► Mixed infections were detected by the multiplexed assay and the AIV H-typing assay. 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 101–103 TCID50/mL and 200–400 EID50/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 EID50/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.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2012.07.005