Volume 10, Issue 40 (8-2020)                   NCMBJ 2020, 10(40): 65-76 | Back to browse issues page

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dehghan esmatabadi M J, barati M, yaghobi moghaddam M A. Design and production of a genetically simulant construct for the detection of B. malei and pseudomaleii. NCMBJ. 2020; 10 (40) :65-76
URL: http://ncmbjpiau.ir/article-1-1316-en.html
Department of Medical Biotechnology, School of Allied Medical Sciences Iran University of Medical Sciences, Tehran, Iran
Abstract:   (672 Views)
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
Material & Methods: 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.
Results: the minimum and maximum sensitivity were 0.1 pg and 10 ng on the constructs designed in the present study, was obtained.
Conclusion: 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.
Full-Text [PDF 681 kb]   (48 Downloads)    
Type of Study: Research Article | Subject: Cellular and molecular
Received: 2020/10/15 | Accepted: 2020/08/31 | Published: 2020/08/31

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