Simultaneous detection and differentiation of Newcastle disease and avian influenza viruses using oligonucleotide microarrays.
Newcastle disease (ND) and avian influenza (AI) are two of the most important zoonotic viral diseases of birds throughout the world. These two viruses often have a great impact upon the poultry industry. Both viruses are associated with transmission from wild to domestic birds, and often display similar signs that need to be differentiated. A rapid surveillance among wild and domestic birds is important for early disease detection and intervention, and is the basis for what measures should be taken. The surveillance, thus, should be able to differentiate the diseases and provide a detailed analysis of the virus strains. Here, we described a fast, simultaneous and inexpensive approach to the detection of Newcastle disease virus (NDV) and avian influenza virus (AIV) using oligonucleotide microarrays. The NDV pathotypes and the AIV haemagglutinin subtypes H5 and H7 were determined at the same time. Different probes on a microarray targeting the same gene were implemented in order to encompass the diversified virus strains or provide multiple confirmations of the genotype. This ensures good sensitivity and specificity among divergent viruses. Twenty-four virus isolates and twenty-four various combinations of the viruses were tested in this study. All viruses were successfully detected and typed. The hybridization results on microarrays were clearly identified with the naked eyes, with no further imaging equipment needed. The results demonstrate that the detection and typing of multiple viruses can be performed simultaneously and easily using oligonucleotide microarrays. The proposed method may provide potential for rapid surveillance and differential diagnosis of these two important zoonoses in both wild and domestic birds.
Wang
Lih-Chiann
LC
Graduate Institute of Veterinary Medicine, National Taiwan University, 1 Sec. 4, Roosevelt Road, Taipei 106, Taiwan.
Pan
Chu-Hsiang
CH
Severinghaus
Lucia Liu
LL
Liu
Lu-Yuan
LY
Chen
Chi-Tsong
CT
Pu
Chang-En
CE
Huang
Dean
D
Lir
Jihn-Tsair
JT
Chin
Shih-Chien
SC
Cheng
Ming-Chu
MC
Lee
Shu-Hwae
SH
Wang
Ching-Ho
CH
D000818:Animals; D001483:Base Sequence; D001717:Birds; D016679:Genome, Viral; D005838:Genotype; D009980:Influenza A virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology; D025202:Molecular Diagnostic Techniques / Q000379:methods / Q000662:veterinary; 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*; D010802:Phylogeny; D011200:Poultry; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D013045:Species Specificity