A novel high-resolution melting analysis-based method for Salmonella genotyping11 Dec 2019 08:54
To establish a simple and rapid high-resolution melting curve (HRM) method, 5 different strains of Salmonella were identified by adding DNA denaturants at different concentrations into the HRM system to change the characteristics of DNA melting and to obtain different Tm (dissolving temperature) values of DNA from different target bacteria. When the concentration of n-butanol was 7% (v/v), the Tm value of the melting curve of the 5 strains changed from 89 °C to 80.5 °C, 81.5 °C, 79.5 °C, 81.0 °C and 82.5 °C, respectively. The sensitivity and specificity of the proposed method were both over 90% in the detection of 270 spiked milk powder samples. In summary, the proposed method in this study has potential for application to food safety and epidemiological research on Salmonella infection.
In the genotyping of Salmonella,
the Tm values of many Salmonella species are basically the same due to
the small genetic differences between species and strains of the same genus,
leading to a failure to distinguish using conventional HRM. In order to expand
the application of HRM in Salmonella genotyping, it is proposed to
modify HRM by screening different DNA denaturants and increase Tm value differences
to obtain a simple method to distinguish Salmonella. This study is of
great significance for the effective prevention and control of Salmonella
infection, and also provides a more rapid and convenient detection method for
related epidemiological studies.