An indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of Type III virulence phenotypes of Pseudomonas aeruginosa isolates.
An indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of Type III virulence phenotypes of Pseudomonas aeruginosa isolates.
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DOI：
10.1186/1476-0711-4-22
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发表时间：
2005-12-23
影响因子：
5.7
通讯作者：
Kazmierczak, Barbara I
中科院分区：
文献类型：
--
作者：
Li, Li;Ledizet, Michel;Kazmierczak, Barbara I
背景：铜绿假单胞菌临床分离株中III型分泌系统的存在与该菌引起的感染的严重疾病和不良结局有关。我们描述了一种间接的酶联免疫吸附试验，该方法可以快速、定量地检测铜绿假单胞菌III型分泌系统的两种外毒素ExoU和ExoT以及两种结构成分PopD和PcrV。将检测结果与标准免疫印迹检测结果进行比较，并与III型分泌依赖性毒力测定结果进行相关性分析。结果：74株分离物中有73株与免疫印迹检测结果一致。结论：建立一种快速检测III型分泌物毒力因子的ELISA法，将有助于开展大型临床研究，以确定铜绿假单胞菌的III型分泌物表型是否能预测患者的临床病程，并在确定感染患者的最佳治疗策略时予以考虑。
BACKGROUND: The presence of a Type III secretion system in clinical isolates of Pseudomonas aeruginosa is associated with severe disease and poor outcomes in infections caused by this pathogen. We describe an indirect enzyme-linked immunosorbent assay that rapidly and quantitatively detects two exotoxins, ExoU and ExoT, and two structural components, PopD and PcrV, of the P. aeruginosa Type III secretion system after in-vitro growth in a calcium-free minimal medium.METHODS: We used this assay to characterize the Type III secretion phenotype of 74 clinical isolates of P. aeruginosa. Findings were compared with results of standard immunoblotting and correlated with Type III secretion-dependent virulence of isolates toward cultured epithelial cells.RESULTS: Results of the ELISA assay were concordant with immunoblot detection of the secreted antigens for 73 of 74 isolates. The Type III secretion phenotype assessed by this immunoassay predicted bacterial virulence toward epithelial cells in vitro for all but five of the clinical isolates.CONCLUSION: The availability of an ELISA assay for rapid detection of Type III secreted virulence factors will facilitate large clinical studies to examine whether the Type III secretion phenotype of a P. aeruginosa isolate predicts the course of clinical disease in a patient and should be taken into account in determining optimal treatment strategies for infected patients.