Department of Genetics and Microbiology, Faculty of Science, Charles University, Prague, Czech Republic.
Lett Appl Microbiol. 2012 Jan;54(1):59-66. doi: 10.1111/j.1472-765X.2011.03172.x. Epub 2011 Nov 23.
The aim of this study was to develop a simple protocol for a PCR-based fingerprinting of Stenotrophomonas maltophilia (SmrepPCR) that utilizes primers complementary to repetitive extragenic palindromic elements (REPs) of this micro-organism.
The relatedness of 34 isolates of environmental and clinical origin was investigated by two SmrepPCRs with two different primers, gyrB sequencing and XbaI macrorestriction followed by pulsed-field gel electrophoresis. While SmrepPCR (with primer DIR) results matched data obtained from the analysis of gyrB nucleotide sequences and identified several clonal complexes, XbaI macrorestriction showed high level of heterogeneity between isolates. The macrorestriction-based clustering of isolates did not correspond to both gyrB and DIR-SmrepPCR grouping.
Our results show that SmrepPCR-inferred relationship of isolates is in a good agreement with sequence-based methods. The combined information from all methods used suggests that rapid evolution of S. maltophilia genomes might be predominantly due to high rate of rearrangements caused by mobile genetic elements.
The presented method is an inexpensive and easy to perform alternative to genotype S. maltophilia isolates and to study their population genetics. SmrepPCR demonstrates the usefulness of species-specific repetitive elements in genomic analyses.
本研究旨在开发一种基于聚合酶链反应(PCR)的简单指纹图谱分析方法,用于 Stenotrophomonas maltophilia(SmrepPCR),该方法利用与该微生物重复外回文元件(REPs)互补的引物。
通过两种不同引物的 SmrepPCR(用引物 DIR)、gyrB 测序和 XbaI 宏观限制后脉冲场凝胶电泳,研究了 34 株环境和临床来源的分离株的相关性。虽然 SmrepPCR(用引物 DIR)的结果与 gyrB 核苷酸序列分析获得的数据相匹配,并鉴定出几个克隆复合体,但 XbaI 宏观限制显示出分离株之间高度的异质性。基于宏观限制的分离株聚类与 gyrB 和 DIR-SmrepPCR 分组均不对应。
我们的结果表明,基于 SmrepPCR 的分离株相关性与基于序列的方法一致。所有方法的综合信息表明,S. maltophilia 基因组的快速进化可能主要是由于移动遗传元件引起的高重组率所致。
本研究提供了一种经济、易于实施的替代方法,用于对 S. maltophilia 分离株进行基因分型,并研究其种群遗传学。SmrepPCR 证明了种特异性重复元件在基因组分析中的有用性。