Establishment and application of a loop-mediated isothermal amplification method for rapid diagnosis of Vibrio cholerae / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To establish a loop-mediated isothermal amplification (LAMP) method for rapid diagnosis of Vibrio cholerae.</p><p><b>METHODS</b>Based on the ompW nucleic sequence of Vibrio cholerae, a pair of primers was designed for LAMP. The reaction conditions were optimized, and the specificity, sensitivity, and practicability of LAMP were tested using 47 bacterial strains and simulated contaminated sites.</p><p><b>RESULTS</b>The results of viable bacterium count showed that LAMP was capable of detecting Vibrio cholerae at a level as low as 1.6x10(2) cfu/ml. The minimal detectable concentration was 1.6+10(3) cfu/ml for simulated contaminated samples such as feces and seawater, and 1.6+10(4) cfu/ml for contaminated milk. All the 21 strains of Vibrio cholerae yielded positive results in LAMP, and the 26 strains of other bacteria all showed negative results, with a detection specificity of 100%.</p><p><b>CONCLUSION</b>The established LAMP method has high specificity and sensitivity for detecting Vibrio cholerae and is applicable in field monitoring and epidemiological study of Vibrio cholerae.</p>