State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China; School of Food Science and Technology, Dalian Polytechnic University, National Engineering Research Center of Seafood, Dalian 116034, China.
Anal Chim Acta. 2017 Mar 22;959:83-90. doi: 10.1016/j.aca.2016.12.045. Epub 2017 Jan 5.
A chemiluminescence resonance energy transfer aptasensor was fabricated for the detection of Staphylococcus aureus (S. aureus) with Co enhanced N-(aminobutyl)-N-(ethylisoluminol) (ABEI) functional flowerlike gold nanoparticles (Co/ABEI-AuNFs) as donor and WS nanosheet as acceptor. In the presence of S. aureus, rolling circle amplification (RCA) can be started. Partially complementary sequence of RCA product functional ABEI-AuNFs (cDNA-ABEI-AuNFs) were then annealed to multiple sites of the RCA product to form duplex complex. This complex is less adsorbed onto the WS nanosheet, thus attenuating the quenching of ABEI-AuNFs chemiluminescence by WS nanosheet. In the absence of target S. aureus (and hence the absence of RCA and duplex formation), the free cDNA-ABEI-AuNFs is completely adsorbed onto the WS nanosheet and chemiluminescence quenching ensues. Under optimal conditions, the logarithmic correlation between the concentration of S. aureus and the CL signal was found to be linear within the range of 50 cfu/mL to 1.5 × 10 cfu/mL (R = 0.9913). The limits of detection of the developed method were found to be 15 cfu/mL for S. aureus. The selectivity and the capability of the biosensor in meat samples were also studied. Therefore, this simple and easy operation method can be used to detect S. aureus with high sensitivity and specificity.
一种化学发光共振能量转移适体传感器被构建用于检测金黄色葡萄球菌(S. aureus),该传感器以 Co 增强的 N-(氨丁基)-N-(乙基异鲁米诺)(ABEI)功能化花状金纳米颗粒(Co/ABEI-AuNFs)作为供体,WS 纳米片作为受体。在存在金黄色葡萄球菌的情况下,可以启动滚环扩增(RCA)。然后,RCA 产物的部分互补序列功能化 ABEI-AuNFs(cDNA-ABEI-AuNFs)退火到 RCA 产物的多个位点以形成双链复合物。该复合物较少地吸附到 WS 纳米片上,从而减弱了 WS 纳米片对 ABEI-AuNFs 化学发光的猝灭。在不存在靶标金黄色葡萄球菌(因此不存在 RCA 和双链形成)的情况下,游离的 cDNA-ABEI-AuNFs 完全吸附到 WS 纳米片上,随后发生化学发光猝灭。在最佳条件下,发现金黄色葡萄球菌浓度与 CL 信号之间的对数相关性在 50 cfu/mL 至 1.5×10 cfu/mL 的范围内呈线性(R=0.9913)。该方法的检测限被发现为金黄色葡萄球菌的 15 cfu/mL。还研究了该生物传感器在肉样中的选择性和检测能力。因此,这种简单易用的操作方法可以用于高灵敏度和特异性地检测金黄色葡萄球菌。