Joung Hyou-Arm, Lee Nae-Rym, Lee Seok Ki, Ahn Junhyoung, Shin Yong Beom, Choi Ho-Suk, Lee Chang-Soo, Kim Sanghyo, Kim Min-Gon
BioNanotechnology Research Center, KRIBB, Daejeon, Republic of Korea.
Anal Chim Acta. 2008 Dec 23;630(2):168-73. doi: 10.1016/j.aca.2008.10.001. Epub 2008 Oct 14.
A signal enhancing method allowing highly sensitive detection of E. coli 16s rRNA was developed using peptide nucleic acid (PNA) as a capture probe and a surface plasmon resonance (SPR) sensor as a detector. 16s rRNA has been used as a genetic marker for identification of organisms, and can be analyzed directly without PCR amplification due to the relatively high number of copies. PNA has a neutral backbone structure, therefore hybridization with 16s rRNA results in the ionic condition being changed from neutral to negative. A cationic Au nanoparticle was synthesized and used for signal amplification by ionic interaction with 16s rRNA hybridized on the PNA probe-immobilized SPR sensor chip. This method resulted in a detection limit of E. coli rRNA of 58.2+/-1.37 pg mL(-1). Using this analytical method, Staphylococcus aureus was detected without purification of rRNA.
开发了一种信号增强方法，该方法使用肽核酸（PNA）作为捕获探针，表面等离子体共振（SPR）传感器作为检测器，可对大肠杆菌16s rRNA进行高灵敏度检测。16s rRNA已被用作生物体鉴定的遗传标记，由于其拷贝数相对较高，无需PCR扩增即可直接进行分析。PNA具有中性主链结构，因此与16s rRNA杂交会导致离子条件从中性变为阴性。合成了阳离子金纳米颗粒，并通过与固定在PNA探针上的SPR传感器芯片上杂交的16s rRNA进行离子相互作用来进行信号放大。该方法对大肠杆菌rRNA的检测限为58.2±1.37 pg mL（-1）。使用这种分析方法，无需纯化rRNA即可检测到金黄色葡萄球菌。
