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Development of a loop-mediated isothermal amplification assay for rapid detection of Burkholderia mallei

Cellular and molecular biology (Noisy-le-Grand, France) | 10 Sep 2016

S Mirzai, S Safi, N Mossavari, D Afshar and M Bolourchian
Abstract
The present study was conducted to establish a Loop-mediated isothermal amplification (LAMP) technique for the rapid detection of B. mallei the etiologic agent of glanders, a highly contagious disease of equines. A set of six specific primers targeting integrase gene cluster were designed for the LAMP test. The reaction was optimized using different temperatures and time intervals. The specificity of the assay was evaluated using DNA from B.pseudomallei and Pseudomonas aeruginosa. The LAMP products were analyzed both visually and under UV light after electrophoresis. The optimized conditions were found to be at 63ÂșC for 60 min. The assay showed high specificity and sensitivity. It was concluded that the established LAMP assay is a rapid, sensitive and practical tool for detection of B. mallei and early diagnosis of glanders.
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Concepts
Infectious disease, Pseudomonas aeruginosa, DNA, Type I and type II errors, Time, Burkholderia, Sensitivity and specificity, Burkholderia mallei
MeSH headings
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