glanders, a biological warfare agent, caused by a bacterium called B mallei. It is considered a contagious, fatal, zoonotic disease. The aim of this study was to establish a fast and reliable molecular method for identification of Burkholderia mallei, this technique was used to determine the Phylogenetic tree and genetic relationship between local and international isolates of B. mallei in horses of Iraq. The research was conducted by taking 100 nasal swabs from animals, and it was found that 29 cases were positive in the PCR test, and 4 random samples were taken from the positive cases for the purpose of sequence analysis.