Colorimetric enzyme-linked aptamer assay utilizing hybridization chain reaction for determination of bovine pregnancy-associated glycoproteins
文献类型： 外文期刊
作者： Lu, Chunxia 1 ; Liu, Changbin 2 ; Shi, Guoqing 2 ;
作者机构： 1.Yangtze Normal Univ, Coll Life Sci & Technol, Chongqing 408100, Peoples R China
2.Xinjiang Acad Agr & Reclamat Sci, Inst Anim Husb & Vet Sci, Shihezi 832000, Peoples R China
关键词： Pregnancy-associated glycoproteins; SELEX; High-throughput sequencing; Colorimetric assay; Pregnancy diagnosis
期刊名称：MICROCHIMICA ACTA （ 2020影响因子：5.833； 五年影响因子：5.357 ）
ISSN： 0026-3672
年卷期： 2020 年 187 卷 6 期
收录情况： SCI
摘要： 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-d = 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. 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
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