Immunology Department, State Research Institute Centre for Innovative Medicine, LT-08409 Vilnius, Lithuania.
Diagn Microbiol Infect Dis. 2011 Nov;71(3):192-200. doi: 10.1016/j.diagmicrobio.2011.06.012.
A quick and robust Salmonella spp. differentiation method based on high-resolution DNA melting (HRM) was developed. DNA samples from 134 Salmonella spp. strains and 20 serotypes were tested. Each serotype was represented by at least 2 strains. All raw data were derived on the Rotor-Gene 65H0-100 system using the designed 8 primer pairs. The reference samples for HRM error evaluation between runs were applied. Raw data error minimization and fluorescence normalization between runs were carried out by application of the proposed calculations. The data analysis showed that repetitive sequence targets are much more informative than the nonrepetitive ones. The method possesses a high potential and can be adopted for further subtyping analyses.
建立了一种基于高分辨率 DNA 熔解（HRM）的快速、稳健的沙门氏菌属区分方法。对来自 134 株沙门氏菌属菌株和 20 个血清型的 DNA 样本进行了测试。每个血清型至少由 2 株菌株代表。所有原始数据均使用设计的 8 对引物在 Rotor-Gene 65H0-100 系统上获得。应用了参考样本进行 HRM 运行间误差评估。通过应用建议的计算方法，实现了运行间原始数据误差最小化和荧光归一化。数据分析表明，重复序列靶标比非重复序列靶标更具信息量。该方法具有很高的潜力，可用于进一步的亚型分析。
