Selective Capture and Identification of Pathogenic Bacteria Using an Immobilized Siderophore
Selective Capture and Identification of Pathogenic Bacteria Using an Immobilized Siderophore
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DOI：
10.1021/la101962w
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发表时间：
2010-10-05
期刊：
影响因子：
3.9
通讯作者：
Low, Philip S.
中科院分区：
文献类型：
--
作者：
Doorneweerd, Derek D.;Henne, Walter A.;Low, Philip S.
快速识别传染性病原体是限制传染病传播的重要一步。虽然基于抗体的检测策略通常因其速度而被选择，但病原体的突变可能会使此类测试过时。为了开发一种快速且防突变的病原体识别方法，我们探索了使用“不可变配体”在检测设备上捕获所需的微生物。在这项“原理验证”研究中，我们将pyoverdine（铜绿假单胞菌必须结合以获得铁）固定到镀金玻璃芯片上，然后检查铁载体捕获铜绿假单胞菌以进行后续鉴定的能力。我们证明，将pyoverdine包被的芯片暴露于不断增加的铜绿假单胞菌稀释度中，可以检测浓度低至10(2)/mL的细菌。我们进一步证明，在检测芯片上以周期性图案打印铁载体，可以通过对图案芯片散射的光进行傅里叶变换分析来实现检测结合病原体的灵敏方法。由于pyoverdine芯片上没有捕获无关的细菌，因此我们得出结论，pyoverdine可用于铜绿假单胞菌的特异性结合和鉴定。由此可见，利用其他微生物特异性的“不可变配体”可以特异性识别其同源病原体。
Rapid identification of infectious pathogens constitutes an important step toward limiting the spread of contagious diseases. Whereas antibody-based detection strategies are often selected because of their speed, mutation of the pathogen can render such tests obsolete. In an effort to develop a rapid yet mutation-proof method for pathogen identification, we have explored the use of "immutable ligands" to capture the desired microbe on a detection device. In this "proof-of-principle" study, we immobilize pyoverdine, a siderophore that Pseudomonas aeruginosa must bind to obtain iron, onto gold-plated glass chips and then examine the siderophore's ability to capture P. aeruginosa for its subsequent identification. We demonstrate that exposure of pyoverdine-coated chips to increasing dilutions of P. aeruginosa allows detection of the bacterium down to concentrations as low as 10(2)/mL. We further demonstrate that printing of the siderophore in a periodic pattern on the detection chip enables a sensitive method of detecting the bound pathogen by a Fourier transform analysis of light scattered by the patterned chip. Because unrelated bacteria are not captured on the pyoverdine chip, we conclude that pyoverdine can be exploited for the specific binding and identification of P. aeruginosa. It follows that the utilization of other microbe-specific "immutable ligands" may allow the specific identification of their cognate pathogens.