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Matteo Perini 1, Gherard Batisti Biffignandi 2, Stella Papaleo 1, Alessandro Alvaro 1, Francesco Comandatore 1 (Pediatric Clinical Research Center “Romeo and Enrica Invernizzi”, Università Di Milano, Milan, Italy; University of Pavia, Pavia, Italia)
Hospital surveillance is one of the most effective strategies to control pathogen spreading in hospital settings. High Resolution Melting (HRM) is an inexpensive PCR-based method to discriminate amplicons on the basis of their melting temperatures and it has the potential to be used for pathogen typing. Despite this, the application of HRM in hospital settings is limited because it is challenging to develop discriminatory typing protocols and to interpret HRM results. Here we present Multi-Locus Melting Clustering (MLMC), a novel approach based on HRM protocols to perform repeatable and portable epidemiological investigations. The MLMC approach includes three main steps: i) the PCR - HRM experiments on the DNA of the isolates, ii) the isolate clustering based on the melting temperatures obtained from the experiments, iii) production of easy-to-read graphical representations using the isolate clusters and the patient/isolates metadata. These graphs help to reconstruct epidemiological scenarios and to identify pathogen transmission routes. MLMC allows to perform fast epidemiological investigations at a fraction of the cost of sequencing-based methods.