Carloni Elisa, Andreoni Francesca, Omiccioli Enrica, Villa Laura, Magnani Mauro, Carattoli Alessandra
Department of Biomolecular Science, University of Urbino "Carlo Bo", via Arco d'Augusto, 2, 61032 Fano, Italy.
Department of Biomolecular Science, University of Urbino "Carlo Bo", via Arco d'Augusto, 2, 61032 Fano, Italy.
Plasmid. 2017 Mar;90:10-14. doi: 10.1016/j.plasmid.2017.01.005. Epub 2017 Jan 27.
Plasmids are the main vectors of resistance and virulence genes in Enterobacteriaceae and plasmid typing is essential for the analysis of evolution, epidemiology and spread of antibacterial resistance. The PCR-Based Replicon Typing (PBRT), developed by Carattoli et al. in 2005, was an efficient method for plasmid identification and typing in Enterobacteriaceae. The 2005 PBRT scheme detected 18 replicons in 8 PCR reactions. Recently, the identification of novel replicons and plasmid types requested an update of the PBRT scheme. A commercial PBRT-KIT was devised for the identification of 28 different replicons in 8 multiplex PCRs. Here we report sensitivity and specificity of the PBRT-KIT carried out in comparison with the 2005 PBRT. The analysis of plasmid content was performed on forty-two enterobacterial strains from different sources, containing different replicon content. The 2005 PBRT identified replicons in 76.2% of the strains. The PBRT-KIT detected replicons in 100% of the analyzed strains, demonstrating increasing sensitivity and specificity of the commercial test with respect to the former 2005 PBRT scheme.
质粒是肠杆菌科细菌中耐药基因和毒力基因的主要载体，质粒分型对于分析抗菌药物耐药性的演变、流行病学和传播至关重要。2005年Carattoli等人开发的基于PCR的复制子分型（PBRT）是一种在肠杆菌科中进行质粒鉴定和分型的有效方法。2005年的PBRT方案在8个PCR反应中检测到18种复制子。最近，新型复制子和质粒类型的鉴定要求更新PBRT方案。设计了一种商业PBRT试剂盒，用于在8个多重PCR中鉴定28种不同的复制子。在此，我们报告了与2005年PBRT相比进行的PBRT试剂盒的敏感性和特异性。对来自不同来源、含有不同复制子含量的42株肠杆菌菌株进行了质粒含量分析。2005年PBRT在76.2%的菌株中鉴定出复制子。PBRT试剂盒在100%的分析菌株中检测到复制子，表明该商业检测相对于以前的2005年PBRT方案具有更高的敏感性和特异性。
