Shi H, Chen Z, Kan J
College of Food Science, Southwest University, Chongqing, China.
Lett Appl Microbiol. 2015 Oct;61(4):361-6. doi: 10.1111/lam.12469.
Pseudomonas aeruginosa is a well-known environmental bacterium capable of causing a variety of life-threatening human infections, with a Type III Secretion System (T3SS) as the most significant virulence determinant. P. aeruginosa strains exhibit unique T3SS virulence genotypes defined by the presence of either exoS or exoU. In this study, loop-mediated isothermal amplification (LAMP) assays for rapid detection of exoS and exoU in P. aeruginosa have been developed and evaluated. Set of four primers were designed for LAMP-based amplification of exoS and exoU respectively. The LAMP reactions were performed at 63°C for 40 min, with detection limits of 100 fg purified DNA. In 107 river water isolates, exoS and exoU were detected in 10 (9%) and 89 (83%) isolates, respectively, and in 38 soil isolates, they were detected in 7 (18%) and 31 (82%) cases respectively. In conclusion, the LAMP assays are rapid, simple and cost-effective tools for detection of the exoU- and exoS-types of P. aeruginosa strains.
This method can be used for the rapid, sensitive and low-cost detection of genes (exoS and exoU) encoding proteins that are part of Type III Secretion System of Pseudomonas aeruginosa. It can serve as an efficient method in outbreak situations or in routine surveillance studies to judge virulence potential and to investigate pathogenesis of P. aeruginosa.
铜绿假单胞菌是一种著名的环境细菌，能够引发多种危及生命的人类感染，其三型分泌系统（T3SS）是最重要的毒力决定因素。铜绿假单胞菌菌株表现出独特的T3SS毒力基因型，由exoS或exoU的存在来定义。在本研究中，已开发并评估了用于快速检测铜绿假单胞菌中exoS和exoU的环介导等温扩增（LAMP）检测方法。分别设计了一组四条引物用于基于LAMP的exoS和exoU扩增。LAMP反应在63°C下进行40分钟，纯化DNA的检测限为100 fg。在107株河水分离株中，分别在10株（9%）和89株（83%）中检测到exoS和exoU，在38株土壤分离株中，分别在7株（18%）和31株（82%）中检测到。总之，LAMP检测方法是用于检测铜绿假单胞菌菌株exoU型和exoS型的快速、简单且经济高效的工具。
该方法可用于快速、灵敏且低成本地检测编码作为铜绿假单胞菌三型分泌系统一部分的蛋白质的基因（exoS和exoU）。它可作为一种有效的方法，在暴发情况或常规监测研究中判断毒力潜力并研究铜绿假单胞菌的发病机制。
