| Info. | Vol.3 - No.1 (2009.03.20) |
|---|---|
| Title | A Diagnostic Microarray for Subtyping and Pathotyping Avian Influenza Virus |
| Authors | Dong-Hun Lee1,†, In-Hyuk Chung2,† Mi-Ra Roh3, Hyun-Jeong Lee1, Youn-Jeong Lee4, Jin-Wook Jung2, Joong-Bok Lee1, Seung-Yong Park1, In-Soo Choi1, Yong-Ho In3, Seung Yong Hwang2 & Chang-Seon Song1 |
| Institutions | 1Avian Disease Laboratory, College of Veterinary Medicine,
Konkuk University, Seoul, Korea 2Department of Biochemistry, Hanyang University & GenoCheck Co., LTD., Sa-Dong, Sangrok-Gu, Ansan, Gyeonggi-Do, Korea 3BIT Center, CT & D Ltd., Seoul, Korea 4Avian Disease Division, National Veterinary Research and Quarantine Service, Ministry for Food, Agriculture, Forestry and Fisheries, Anyang, Korea †These authors contributed equally to this work Correspondence and requests for materials should be addressed to C.S. Song (songcs@konkuk.ac.kr) |
| Abstract | The ability to readily identify new and potentially pandemic strains of influenza virus will allow a more rapid response by health care officials to reduce the spread and human impact of the disease. We report a low-density microarray for the rapid detection and identification of potential pandemic influenza A virus subtypes H5, H7, and H9 in the present study. Subtype- and pathotype-specific probes were designed to target the hemagglutinin (HA) gene segment. The system consisted of RT-PCR using universal primers for the HA gene. RT-PCR products were hybridized against a microarray consisting of short probes 31- 41 nucleotides in length. Cy3-labeled DNA targets were generated with a Cy3-labeled primer. Reference strains (n=46) of different avian influenza subtypes (H1-H15, except H16), and 34 unidentified field strains of avian influenza virus (AIV) from wild bird habitats and live bird markets was used for validation. All referenced subtypes belonged to H5, H7, and H9 and unidentified field strains were correctly typed by their HA-subtypes and pathotypes. These included 3 highly pathogenic H5N1 avian influenza viruses which had caused South Korean outbreaks in 2003, 2005, and 2008. In addition, there were no cross-hybridization reactions with other poultry espiratory viruses, suggesting that the microarray was specific for influenza A iruses. The developed microarray was capable of subtyping and pathotyping the otential pandemic influenza A virus subtypes H5, H7, and H9. |
| Keyword | Avian influenza, Microarray analysis, Hemagglutinin |
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