Development of a multiple-run high-resolution melting assay for Salmonella spp. genotyping HRM application for Salmonella spp. subtyping

Maksim Bratchikov; Mykolas Mauricas

doi:10.1016/j.diagmicrobio.2011.06.012

Development of a multiple-run high-resolution melting assay for Salmonella spp. genotyping HRM application for Salmonella spp. subtyping

Diagn Microbiol Infect Dis. 2011 Nov;71(3):192-200. doi: 10.1016/j.diagmicrobio.2011.06.012.

Authors

Maksim Bratchikov¹, Mykolas Mauricas

Affiliation

¹ Immunology Department, State Research Institute Centre for Innovative Medicine, LT-08409 Vilnius, Lithuania. maksim@pcr.lt

PMID: 21996359
DOI: 10.1016/j.diagmicrobio.2011.06.012

Abstract

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.

MeSH terms

Bacterial Typing Techniques*
Genotyping Techniques*
Real-Time Polymerase Chain Reaction*
Reproducibility of Results
Salmonella / classification*
Salmonella / genetics
Sensitivity and Specificity