The PBRT (PCR Based Replicon Typing) is an essential assay for the analysis of evolution, epidemiology and spread of antibacterial resistance. The exclusive assay allows fast and easy plasmid characterization and molecular identification through replicons detection in Enterobacteriaceae. The PBRT 2.0 kit is the updated version of the PBRT-KIT including all the modifications added to the 2005 PBRT (Carattoli et al. 2005) along the years and new epidemiologically relevant replicons P1, N2, FIB-KN, FIB-KQ, X4. The new updated version strongly improves the detection of plasmid content and replicon diversity with respect to previous version of PBRT scheme exhibiting a good specificity and sensitivity. PBRT 2.0 kit provides PCR assays and positive controls to perform the amplification of 30 replicons (HI1, HI2, HIB-M, I1 a, I1 g, I2, X1, X2, X3, X4, BO, M, N, N2, FII, FIA, FIB, FIB-KQ, FIB KN, FIB-M, FIIK , FIIS, P1, R, U, L, A/C, T, W).