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Detection and Identification of Avian Influenza Virus by cDNA Microarray

Michele N Maughan, Travis W Bliss, Ida Chung, David L Suarez and Calvin L Keeler Jr

Influenza A viruses consisting of all known 16 HA and 9 NA subtypes have been isolated from birds. We have created a diagnostic avian cDNA microarray containing probes corresponding to the highly conserved matrix (M) gene, and selected hemagglutinin (HA), and neuraminidase (NA) subtypes of AIV. cDNA RT-PCR products from the HA, NA, and M genes of various avian influenza isolates and subtypes were used to create an avian influenza virus (AIV) cDNA microarray. The microarray was evaluated against a panel of AIV isolates in order to appraise its application in AIV detection and identification. Utilizing the M gene as a pan-influenza marker, all 10 samples were identified as being strains of type A influenza. The array was able to correctly HA- and NA-subtype subtype 7 out of 10 test samples. This included correctly identifying, subtyping, and determining the geographic origin of all of the H5 subtypes and the two H7 samples of U.S. origin.