Wen Junlin, Zhou Shungui, Chen Junhua
Guangdong Institute of Eco-environmental and Soil Sciences, Guangzhou 510650, P. R. China.
Sci Rep. 2014 Jun 5;4:5191. doi: 10.1038/srep05191.
Rapid detection and enumeration of target microorganisms is considered as a powerful tool for monitoring bioremediation process that typically involves cleaning up polluted environments with functional microbes. A novel colorimetric assay is presented based on immunomagnetic capture and bacterial intrinsic peroxidase activity for rapidly detecting Shewanella oneidensis, an important model organism for environmental bioremediation because of its remarkably diverse respiratory abilities. Analyte bacteria captured on the immunomagnetic beads provided a bacterial out-membrane peroxidase-amplified colorimetric readout of the immunorecognition event by oxidizing 3, 3', 5, 5'-tetramethylbenzidine (TMB) in the present of hydrogen peroxide. The high-efficiency of immunomagnetic capture and signal amplification of peroxidase activity offers an excellent detection performance with a wide dynamic range between 5.0 × 10(3) and 5.0 × 10(6) CFU/mL toward target cells. Furthermore, this method was demonstrated to be feasible in detecting S. oneidensis cells spiked in environmental samples. The proposed colorimetric assay shows promising environmental applications for rapid detection of target microorganisms.
快速检测和计数目标微生物被认为是监测生物修复过程的有力工具,生物修复过程通常涉及用功能性微生物清理受污染的环境。本文提出了一种基于免疫磁捕获和细菌固有过氧化物酶活性的新型比色测定法,用于快速检测希瓦氏菌,它是环境生物修复的重要模式生物,因其具有显著多样的呼吸能力。捕获在免疫磁珠上的分析物细菌通过在过氧化氢存在下氧化3,3',5,5'-四甲基联苯胺(TMB),提供了免疫识别事件的细菌外膜过氧化物酶放大比色读数。免疫磁捕获的高效性和过氧化物酶活性的信号放大提供了出色的检测性能,对目标细胞的动态范围在5.0×10(3)至5.0×10(6) CFU/mL之间。此外,该方法在检测环境样品中加标的希瓦氏菌细胞方面被证明是可行的。所提出的比色测定法在快速检测目标微生物方面显示出有前景的环境应用。