Journal
TALANTA
Volume 191, Issue -, Pages 54-58Publisher
ELSEVIER
DOI: 10.1016/j.talanta.2018.08.033
Keywords
Morganella morganii; Isothermal amplification; Endpoint turbidity analysis; Point-of-care diagnostic kit; Infectious disease
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Funding
- Iranian National Science Foundation (INSF), Iran [95849799]
- Shahid Beheshti University
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A point-of-care diagnostic kit was developed for detection of Morganella morganii using optimized loop-mediated isothermal amplification (LAMP) technique within less than an hour. In that regard, dimethyl sulfoxide (DMSO) was utilized together with betaine and all variables were optimized to improve the efficiency of the method. Moreover, surface presentation of antigens protein was targeted and six unique primers were designed. Endpoint turbidity analysis was performed at 550 nm to measure the tetravalent anion (pyrophosphate) released during the reaction. The specificity of the method was evaluated using nine closely related bacterial species as well as its sensitivity. It was shown that the improved LAMP assay could significantly distinguish M. morganii from other bacteria while the sensitivity was determined to be 0.2 CFU mL(-1).
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