Use of a Rose Bengal test for diagnosis of Glanders in equines

Glanders is an infectious disease that is caused by the bacterium Burkholderia mallei. Glanders occurs primarily in equines, but other species, including humans, may become incidental hosts. The allergic test of mallein is a valid and usual test in Iran which is used for the diagnosis of Glanders. The mallein test requires an experienced laboratory person and lasts 48 hours. Therefore in order to quickly diagnose the disease, especially in borders of country, where animals cannot be kept for longtime, new method should be used to identify the disease. To improve identification, in terms of sensitivity and specificity, new methods are needed to identify the disease. The aim of this study is designing of Rose Bengal test (RBT) for diagnosis of Glanders. Sera from 70 naturally infected culture-positive, 3 equines that were sensitized by injecting antigen and 110 healthy equines were tested. Burkholderia mallei strain to produce mallein was cultured in glycerinated nutrient medium. To ensure the presence of the bacterium and determine the identity of the PCR were performed. Antigen for RBT was prepared from Burkholderia mallei and serums were added to Rose bengal antigen. Finally positive and negative serums were analyzed for confirming the accuracy of the test. By performing PCR, the bacteria characteristic band was observed in the range of 250 bp. By examining positive and negative serums using RBT, positive or negative answer was obtained explicitly. By designing and performing this test, it can be consider, this test has been using for Rapid detection of Glanders with features such as, ease of use and can be applicable without specialized equipment and trained personnel.