Carattoli Alessandra, Bertini Alessia, Villa Laura, Falbo Vincenzo, Hopkins Katie L, Threlfall E John
Department of Infectious, Parasitic and Immune-mediated Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy.
J Microbiol Methods. 2005 Dec;63(3):219-28. doi: 10.1016/j.mimet.2005.03.018. Epub 2005 Jun 2.
The epidemiological importance of tracing plasmids conferring drug resistance prompted us to develop a PCR method based on replicons (inc/rep PCR) of the major plasmid incompatibility groups among Enterobacteriaceae. Eighteen pairs of primers were designed to perform 5 multiplex- and 3 simplex-PCRs, recognizing FIA, FIB, FIC, HI1, HI2, I1-Igamma, L/M, N, P, W, T, A/C, K, B/O, X, Y, F, and FIIA. The specificity of the method was tested on a collection of 61 reference plasmids and on 20 Salmonella enterica strains of different serotypes isolated in Italy. Results indicated that the inc/rep PCR method demonstrates high specificity and sensitivity in detecting replicons on reference plasmids and also revealed the presence of recurrent and common plasmids in epidemiologically unrelated Salmonella isolates of different serotypes. These results suggest that the method is potentially applicable to a large number of strains to trace the diffusion of specific multi-drug resistance plasmids in different environments.
追踪赋予耐药性的质粒在流行病学上的重要性促使我们开发一种基于肠杆菌科主要质粒不相容群复制子（inc/rep PCR）的PCR方法。设计了18对引物来进行5次多重PCR和3次单重PCR，可识别FIA、FIB、FIC、HI1、HI2、I1-Iγ、L/M、N、P、W、T、A/C、K、B/O、X、Y、F和FIIA。该方法的特异性在61个参考质粒以及从意大利分离出的20株不同血清型的肠炎沙门氏菌菌株上进行了测试。结果表明，inc/rep PCR方法在检测参考质粒上的复制子时显示出高特异性和敏感性，并且还揭示了不同血清型、在流行病学上不相关的沙门氏菌分离株中存在常见质粒。这些结果表明，该方法可能适用于大量菌株，以追踪特定多药耐药质粒在不同环境中的传播。
