Abstract: Background: Avian influenza is an infectious viral disease of birds which can spread to domestic poultry (chicken, duck and quail) and cause large-scale outbreaks of serious disease. Avian Influenza Virus (AIV) has also been reported to cross the species barrier and cause disease or subclinical infections in other mammals and humans. Based on the antigenic differences in nucleoprotein (NP) and Matrix (M) proteins, AIV is classified into A, B and C types. All influenza viruses infected and isolated from avian are classified as A type. Further subtyping of AIV is based on antigenic differences between the two surface glycoproteins hemagglutinin (H) and neuraminidase (N). Objective: The aim of this study was to identify AIV type A, subtype of H5 by conducting the serological and molecular analysis toward different poultry species which were recently infected by AIV. Materials and Methods: All of 37 samples collected from tracheal, cloacal swabs and visceral organs from various species of poultry (chickens, ducks and quails) were used in this study. The samples were inoculated into Specific Antibody Negative (SAN) embryonated chicken eggs of 9-11 days of age for serological and molecular analysis. For serological analysis, all 37 samples were tested by Hemagglutination Activity (HA) and Hemagglutination Inhibition (HI) tests. For molecular analysis of Matrix (M) and Hemaglutinin 5 (H5) genes, the RNA viruses were extracted and amplified by RT-PCR method. The amplification products were then separated on 1% agarose gel electrophoresis and sybrsafe staining. Results: Serological analysis by HA and HI tests shown that 7 samples from total of 37 samples were positively infected by AIV type A. Molecular analysis by RT-PCR amplification to amplify matrix and hemaglutinin genes followed by separation of RT-PCR products on 1% agarose gel electrophoresis were obtained DNA fragments in size of 276 and 1.725 bp, respectively in 7 tested samples. Conclusion: It could be concluded that the infectious agent which cause of the death of various species of poultry in Aceh province, Indonesia were AIV type A, subtype H5.