Xu Yanyang, Huo Bingyang, Sun Xuan, Ning Baoan, Peng Yuan, Bai Jialei, Gao Zhixian
College of Food Science and Engineering, Jilin University Changchun 130022 P. R. China
Huazhong Agricultural University, College of Life Science and Technology Wuhan 430070 P. R. China.
RSC Adv. 2018 Apr 30;8(29):16024-16031. doi: 10.1039/c8ra01599f. eCollection 2018 Apr 27.
A rapid, simple, and sensitive method has been developed to detect staphylococcal enterotoxin B (SEB). To establish the hybridization chain reaction-based aptasensor, we described the new probes of two hairpins (H1 and H2), which were first designed based on the partial complementary sequence of the SEB aptamer (cDNA). The H1 labeled with a fluorophore and a quencher can act as a molecular fluorescence "switch". Hence, in the presence of SEB, the aptamer binds SEB, while the unbound cDNA triggers HCR to carry out the cyclic hybridization of H1 and H2 so as to turn "ON" the fluorescence through forming long nicked DNA. By using this new strategy, SEB can be sensitively detected within the range of 3.13 ng mL to 100 ng mL with a detection limit of 0.33 ng mL (S/N = 3). Furthermore, the developed method could facilitate the detection of SEB effectively in milk samples.
已开发出一种快速、简单且灵敏的方法来检测葡萄球菌肠毒素B（SEB）。为建立基于杂交链式反应的适体传感器，我们描述了两个发夹结构（H1和H2）的新型探针，它们首先基于SEB适体（cDNA）的部分互补序列设计。标记有荧光团和猝灭剂的H1可作为分子荧光“开关”。因此，在存在SEB的情况下，适体与SEB结合，而未结合的cDNA触发HCR以进行H1和H2的循环杂交，从而通过形成长切口DNA使荧光“开启”。通过使用这种新策略，可在3.13 ng/mL至100 ng/mL范围内灵敏地检测SEB，检测限为0.33 ng/mL（S/N = 3）。此外，所开发的方法能够有效地促进对牛奶样品中SEB 的检测。
