Kim Insu, Kim Yonghwan, Lee Sang Won, Lee Dongtak, Jung Hyo Gi, Jang Jae Won, Lee Taeha, Yoon Young Kyung, Lee Gyudo, Yoon Dae Sung
School of Biomedical Engineering, Korea University, Seoul 02841, South Korea.
School of Biomedical Engineering, Korea University, Seoul 02841, South Korea; Interdisciplinary Program in Precision Public Health, Korea University, Seoul 02841, South Korea.
Biosens Bioelectron. 2021 Aug 1;185:113267. doi: 10.1016/j.bios.2021.113267. Epub 2021 Apr 23.
Without appropriate treatment, Staphylococcus aureus (S. aureus) infection can cause life-threatening diseases (e.g., meningitis, pneumonia, bacteremia, and sepsis). However, a rapid and accurate point-of-care test for the infection remains challenging. The bacterium secretes α-hemolysin (Hla), which spontaneously binds to the cell membrane of erythrocyte, and eventually lyses the cell via pore formation. Taking advantage of this phenomenon, we apply the erythrocyte membrane (EM) extracted from human whole blood as a novel bioreceptor for detecting Hla, fabricating erythrocyte-camouflaged biosensors (ECB) by coating EM onto electrochemical impedance electrodes. We verify the existence of EM on the ECB by using confocal microscopy and atomic force microscopy. We demonstrate that ECBs sensitively detect Hla spiked in phosphate buffer saline and human serum. Also, the sensor shows higher sensitivity to Hla than major blood proteins, such as human serum albumin, fibrinogen, and gamma globulin. Specifically, the signal intensities for Hla are 8.8-12.7 times higher than those in the same concentration of those blood proteins. The detection limit of the ECB for Hla is 1.9 ng/ml while the dynamic range is 0.0001-1 mg/ml. Finally, we validate the constant sensing performance of ECB with 99.0 ± 5.6% accuracy for 35 days of storage.
如果不进行适当治疗,金黄色葡萄球菌(S. aureus)感染可导致危及生命的疾病(如脑膜炎、肺炎、菌血症和败血症)。然而,针对该感染的快速准确的即时检测仍然具有挑战性。这种细菌会分泌α-溶血素(Hla),它会自发地与红细胞细胞膜结合,并最终通过形成孔道使细胞裂解。利用这一现象,我们将从人全血中提取的红细胞膜(EM)用作检测Hla的新型生物受体,通过将EM涂覆在电化学阻抗电极上制备红细胞伪装生物传感器(ECB)。我们使用共聚焦显微镜和原子力显微镜验证了ECB上EM的存在。我们证明ECB能够灵敏地检测添加到磷酸盐缓冲盐水和人血清中的Hla。此外,该传感器对Hla的灵敏度高于主要血液蛋白,如人血清白蛋白、纤维蛋白原和γ球蛋白。具体而言,Hla的信号强度比相同浓度的那些血液蛋白的信号强度高8.8 - 12.7倍。ECB对Hla的检测限为1.9 ng/ml,动态范围为0.0001 - 1 mg/ml。最后,我们验证了ECB在储存35天期间具有99.0 ± 5.6%的准确率,传感性能稳定。