College of Life Science and Technology, Yangtze Normal University, Chongqing, 408100, People's Republic of China.
Institute of Animal Husbandry and Veterinary Science, Xinjiang Academy of Agriculture and Reclamation Science, Shihezi, 832000, People's Republic of China.
Mikrochim Acta. 2020 May 8;187(6):316. doi: 10.1007/s00604-020-04301-y.
DNA aptamers that bind to bovine pregnancy-associated glycoproteins (bPAGs) were selected by the systematic evolution of ligands by exponential enrichment (SELEX) procedure coupled to surface plasmon resonance (SPR) and high-throughput sequencing (HTS) technology. After seven rounds of selection using carboxylated magnetic beads (MB) coated with bovine pregnancy-associated glycoproteins 9 (bPAG9) and bovine serum albumin (BSA) as target and counter targets, respectively, two aptamers designated as A1 and A24 showed high affinities to bPAG9 (K = 1.04 and 2.5 nM). Moreover, the specificity was determined by testing the non-targets bPAG4, bPAG6, bPAG16, BSA, and ovalbumin (OVA). Results showed that two aptamers demonstrated broad group specificity to bPAG family. Subsequently, a colorimetric sandwich enzyme-linked aptamer assay was developed for ultrasensitive detection of bPAG9 based on hybridization chain reaction (HCR) amplification strategy. The method exhibited a broad determination from 0.134 to 134 ng/mL with a detection limit of 0.037 ng/mL. The method has been successfully applied to determine bPAGs in real samples. The results demonstrate that the developed aptamers could be used as promising molecular probes for the development of pregnancy diagnostic tools. Graphical abstract In this study, we first selected aptamers against bovine pregnancy-associated glycoproteins (bPAGs) as molecular recognition elements and then developed a colorimetric enzyme-linked aptamer assay utilizing hybridization chain reaction to detect bPAGs in the serum.The GA can't be deleted, the modified GA can not upload. So themodified GA and figures will be send to CorrAdmin3@spi-global.com.
通过与表面等离子体共振（SPR）和高通量测序（HTS）技术相结合的配体指数富集系统进化（SELEX）程序，筛选出与牛妊娠相关糖蛋白（bPAG）结合的 DNA 适体。经过七轮以牛妊娠相关糖蛋白 9（bPAG9）和牛血清白蛋白（BSA）分别作为靶标和对照靶标包被的羧基化磁珠（MB）为筛选的筛选，得到了两个与 bPAG9 具有高亲和力的适体，分别命名为 A1 和 A24（Kd=1.04 和 2.5 nM）。此外，通过测试非靶标 bPAG4、bPAG6、bPAG16、BSA 和卵清蛋白（OVA）来确定特异性。结果表明，两个适体对 bPAG 家族具有广泛的群体特异性。随后，基于杂交链式反应（HCR）扩增策略，建立了用于超灵敏检测 bPAG9 的比色夹心酶联适体测定法。该方法的测定范围为 0.134 至 134 ng/mL，检测限为 0.037 ng/mL。该方法已成功应用于实际样品中 bPAGs 的测定。结果表明，所开发的适体可作为开发妊娠诊断工具的有前途的分子探针。
