To assess the potency of the PPD-mallein produced in Brazil, five animals were from a property identified as a focus of glanders. These animals had suggestive clinical signs of the disease and the other five, from a property free from glanders, showed no clinical signs and were serology negative (control group). PPD-mallein from Burkholderia mallei was obtained by precipitation with trichloroacetic acid and ammonium sulfate. The animals were inoculated according to the criteria established by Department of Agriculture, Livestock and Supply (MAPA) for the diagnosis of glanders. After 48 h of application of PPD-mallein, there was swelling in the area of application, presence of ocular secretion and tears in sick animals. The control group showed no inflammatory reaction at the site of inoculation of PPD-mallein. This immunogen produced in Brazil and still being tested was effective for identifying the infection in true positive animals and excluding the truly negative ones, being a new possibility for diagnosis and control of glanders.