Castañeda Nancy Claudia, Pichel Mariana, Orman Betina, Binsztein Norma, Roy Paul H, Centrón Daniela
Departamento de Bacteriología, Instituto Nacional de Enfermedades Infecciosas, ANLIS Dr. Carlos G. Malbrán, Av Velez Sarfield 563, 1281, Buenos Aires, Argentina.
Diagn Microbiol Infect Dis. 2005 Nov;53(3):175-83. doi: 10.1016/j.diagmicrobio.2005.05.008. Epub 2005 Oct 24.
We have developed a novel typing method based on Vibrio cholerae repeat sequences (VCR) using primers directed out of the VCR sequences. To evaluate the VCR-polymerase chain reaction (PCR) as a typing system, 2 categories, efficacy and efficiency, were analyzed in 69 strains of human and environmental V. cholerae O1 toxigenic and nontoxigenic, and non-O1 strains isolated since 1992-2000 from Argentina. The discriminatory power (0.91), stability (0.95), reproducibility (1), typeability (1), rapidity, accessibility, as well ease of use, indicated that the VCR-PCR method provides an alternative useful tool for molecular epidemiology of V. cholerae. The VCR-PCR of V. cholerae isolates showed 29 patterns, of which pattern 1 represented 68% of the V. cholerae O1 isolates, supporting the hypothesis that a clone with epidemic behavior was responsible for the epidemic in Latin America. These results showed a good correlation and a better epidemiologic analysis when the results were compared in parallel with repetitive extragenic palindromic sequences-PCR. In conclusion, VCR-PCR showed excellent performance as a typing method for cholera surveillance programs.
我们开发了一种基于霍乱弧菌重复序列(VCR)的新型分型方法,使用的引物指向VCR序列之外。为了评估VCR聚合酶链反应(PCR)作为一种分型系统,我们对1992年至2000年从阿根廷分离出的69株人类和环境来源的霍乱弧菌O1产毒株和非产毒株以及非O1菌株,从效能和效率两个类别进行了分析。其鉴别力(0.91)、稳定性(0.95)、可重复性(1)、分型能力(1)、快速性、可及性以及易用性表明,VCR-PCR方法为霍乱弧菌分子流行病学提供了一种有用的替代工具。霍乱弧菌分离株的VCR-PCR显示出29种模式,其中模式1占霍乱弧菌O1分离株的68%,支持了一种具有流行特性的克隆导致拉丁美洲疫情的假说。当将结果与重复基因外回文序列-PCR平行比较时,这些结果显示出良好的相关性和更好的流行病学分析效果。总之,VCR-PCR作为霍乱监测项目的分型方法表现出色。