Use of Dot-blot technique by Burkholderia mallei for the serodiagnosis of glanders in Iran

Glanders is one of the most important zoonotic diseases that caused by Burkholderia mallei. Considering that Iran as the major center for glanders and risks of transmission of it, quick detection is very important. Therefore, the aim of this study is designing of Dot-Elisa system. So, 40 serum samples were collected from different provinces, along with antigen sample of Burkholderia mallei as positive control and healthy horses’ serum (negative control) was used. The optimum concentrations of antigen-antibody were determined by checkerboard titration using glanders positive and negative horse sera. After designing the dot-blot test, an antigen was coated into the nitrocellulose membrane wells and stopped by blocking solution and the suspicious equine serum was poured into the wells along with positive and negative control. In the following conjugated antibody was added on it, finally substrate was added to the wells and reaction was read as color signal. In this study, out of 40 samples, 6 positive cases were positive. Considering mallein test as a standard test and comparing with DotElisa, the sensitivity and specificity were 100% and 91.9% respectively. The results showed that dot-blot test was successful for detection of Glanders due to its high sensitivity, speed, readability and field applicability.