CAJ | 학술논문

运用纳米金复合探针结合蛋白芯片,建立了一种检测心肌损伤标志物的新方法。构建了2种纳米金探针：标记有检测抗体和DNA探针1的检测探针和标记有DNA探针2(与DNA探针1的碱基互补配对)的信号探针。当目的抗原存在时,检测探针经检测抗体和抗原,与芯片上捕获抗体结合固定在芯片上,信号探针通过碱基互补配对与检测探针结合使信号放大,最后利用纳米金成核原理染色,通过显微镜观察结果并定量分析。该体系在40 min内可检测多种标志物,其中肌钙蛋白Ⅰ( cTnⅠ)的检出限为10 pg/mL,与临床电化学发光法( ECLIA)灵敏度相当；肌红蛋白( MYO)与新型脂肪酸结合蛋白( HFABP)的检出限分别为640和10 pg/mL,与ECLIA及酶联免疫吸附法( ELISA)相比,灵敏度显著提高。
운용납미금복합탐침결합단백심편,건립료일충검측심기손상표지물적신방법。구건료2충납미금탐침：표기유검측항체화DNA탐침1적검측탐침화표기유DNA탐침2(여DNA탐침1적감기호보배대)적신호탐침。당목적항원존재시,검측탐침경검측항체화항원,여심편상포획항체결합고정재심편상,신호탐침통과감기호보배대여검측탐침결합사신호방대,최후이용납미금성핵원리염색,통과현미경관찰결과병정량분석。해체계재40 min내가검측다충표지물,기중기개단백Ⅰ( cTnⅠ)적검출한위10 pg/mL,여림상전화학발광법( ECLIA)령민도상당；기홍단백( MYO)여신형지방산결합단백( HFABP)적검출한분별위640화10 pg/mL,여ECLIA급매련면역흡부법( ELISA)상비,령민도현저제고。
A novel method based on gold nanoparticles( AuNPs) probes and protein chip was developed to de-tect myocardial injury markers. Two AuNPs probes were constructed:one was modified with detection antibody and DNA probe 1(detection probe), and the other was labeled with DNA probe 2 which was complementary to DNA probe 1(signal probe). When target antigen was present, a sandwich structure(capture antibody-target antigen-detection probe-signal probe) was formed through the interaction of the antibody-antigen and the hy-bridization of complemenatry DNAs. Then the chip was dyed by the gold deposition solution, and the signal was amplified. The results were observed with a microscope linked with an image analysis software. This sys-tem could simultaneously detect multiple markers in 40 min. The detection limit of cTnⅠ was 10 pg/mL, which was equivalent to ECLIA in clinical. For HFABP and MYO, the detection limit could be as low as 640 and 10 pg/mL, respectively, and the sensitivity was improved greatly when compared with ECLIA and ELISA.