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Abstract
Asian-lineage H5 highly pathogenic avian influenza (HPAI) viruses have caused continuous outbreaks in poultry and wild birds. Development of rapid and accurate diagnostic methods is needed for preventing further spread of the virus and reducing the time required for eradication of the virus. We developed a low-density microarray for the rapid detection and identification of avian influenza virus subtypes H5, H7, and H9 and their pathotypes in a previous study. In the present study, we evaluated previously developed diagnostic microarray using avian influenza viruses isolated in Mongolia, including H5 HPAI viruses. All H5 HPAI viruses isolated in Mongolia were shown as H5-specific and highly pathogenic pattern in the microarray. H2, H3 and H12 viruses isolated in Mongolia used in this study did not show any H5, H7 and H9 patterns. These results indicated that this diagnostic microarray has enormous potential for the rapid subtyping and pathotyping of influenza viruses, including viruses isolated in Mongolia.
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Figure 1.
Hybridization patterns of influenza viruses isolated in Mongolia. The microarray layout of 69 HA probes, 1 M gene probes (red circles), and position markers (green circles) is indicated. Among the 69 HA probes, there were 19 probes for the H5 subtype (blue circles), 6 probes for the H7 subtype (pink circles), 15 probes for the H9 subtype (sky blue circles), 26 probes for high pathogenicity (yellow circles) and 3 probes for low pathogenicity (purple circles) which were spotted in duplicate on silylated slides (A). Cy3-tagged RT-PCR products were denatured and mixed with hybridization buffer. The mixture was hybridized onto the oligonucleotide arrays for 1 h at 55°C. The oligonucleotide arrays were washed and hybridization signals were detected using a microarray scanner (B).
Table 1.
Influenza viruses isolated from Mongolia used in the study
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