Design and production of a genetically simulant construct for the detection of B. malei and pseudomaleii

Aim & Background:

 the genus Burkholderia (mallei and pseudomallei species) are classified as dangerous human pathogens. Diagnosing cultures for this bacterium is costly and time consuming and on the other hand it is very dangerous to work with and it is very difficult to access this bacterium. the porpose of this research Design and production of a genetically simulant construct for the detection of  B. pseudomaleii

initially, a simulant construct containing the reference and specific genes of the mallei and pseudomallei species was not observed in other species of the family, and then the operational efficiency of the designed construct was quantified using Real-Time Quantitative PCR. Quality (SYBER Green) and qualitative (using TaqMan probe) were evaluated. Finally, Real-Time PCR amplification products were analyzed by melting curve and electrophoresis.

the minimum and maximum sensitivity were 0.1 pg and 10 ng on the constructs designed in the present study, was obtained.

This simulat can be used as a positive control for the detection of the Burkholderia agent and can further enhance the development of new diagnostic kits in the field.