A Colorimetric Detection of bPAG9 Based on Aptamer and Peroxidase-like Mimics of Metal Organic Framework

LU Chun-xia; LIU Chang-bin; ZHANG Zhen-liang; LU Shou-liang; WU Shi-jia

doi:10.12452/j.fxcsxb.24010301

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A Colorimetric Detection of bPAG9 Based on Aptamer and Peroxidase-like Mimics of Metal Organic Framework

Scientific Papers | 更新时间：2024-08-16

- A Colorimetric Detection of bPAG9 Based on Aptamer and Peroxidase-like Mimics of Metal Organic Framework
- Journal of Instrumental Analysis Vol. 43, Issue 5, Pages: 714-721(2024)
- 作者机构：
  
  1.长江师范学院现代农业与生物工程学院，重庆 408100
  2.新疆农垦科学院省部共建绵羊遗传改良与健康养殖国家重点实验室，新疆石河子 832000
  3.江南大学　食品学院，江苏无锡 214122
- 作者简介：
- 基金信息：
- DOI：10.12452/j.fxcsxb.24010301
  CLC： O629.7;S826
- Published：15 May 2024，
  
  Received：03 January 2024，
  
  Revised：18 February 2024，
- 稿件说明：
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卢春霞,刘长彬,张振良,卢守亮,吴世嘉.基于适配体-MOF模拟酶比色检测bPAG9[J].分析测试学报,2024,43(05):714-721.

LU Chun-xia,LIU Chang-bin,ZHANG Zhen-liang,LU Shou-liang,WU Shi-jia.A Colorimetric Detection of bPAG9 Based on Aptamer and Peroxidase-like Mimics of Metal Organic Framework[J].Journal of Instrumental Analysis,2024,43(05):714-721.
卢春霞,刘长彬,张振良,卢守亮,吴世嘉.基于适配体-MOF模拟酶比色检测bPAG9[J].分析测试学报,2024,43(05):714-721. DOI： 10.12452/j.fxcsxb.24010301.

LU Chun-xia,LIU Chang-bin,ZHANG Zhen-liang,LU Shou-liang,WU Shi-jia.A Colorimetric Detection of bPAG9 Based on Aptamer and Peroxidase-like Mimics of Metal Organic Framework[J].Journal of Instrumental Analysis,2024,43(05):714-721. DOI： 10.12452/j.fxcsxb.24010301.

摘要

该研究基于金属有机框架Fe-MIL-101和双适配体夹心原理建立了一种比色检测牛妊娠相关糖蛋白9（bPAG9）的新方法。将生物素化适配体1固定在微孔板上作为捕获探针，Fe-MIL-101功能化的适配体2作为信号探针，当测试样品中含有bPAG9时，适配体1和适配体2与bPAG9结合，在固相载体表面形成双适配体夹心结构，随后适配体2偶联的Fe-MIL-101催化无色过氧化物酶底物产生比色输出信号，通过比色信号的变化即可实现对bPAG9的定性和定量分析。结果显示：在优化条件下，bPAG9在缓冲液中的线性检测范围为0.5～50 ng/mL（

=0.996），检出限为0.13 ng/mL。bPAG9在空白血清中的加标回收率为89.4%~104%，相对标准偏差小于10%。将该方法应用于人工受精28 d的奶牛血清检测，妊娠诊断结果显示，该法的诊断敏感性、特异性和准确率分别为92.5%、90.0%和91.5%。所建方法具有无创、简便、经济等特点，在奶牛早孕检测领域具有良好的应用前景。

Abstract

The aim of this study is to establish a new colorimetric method for the detection of bovine pregnancy associated glycoprotein 9（bPAG9） based on the metal-organic framework Fe-MIL-101 and double aptamer sandwich principle. In a typical experimental procedure，biotinylated aptamer1 was immobilized onto microplate to act as capture probes. Fe-MIL-101 functionalized aptamer 2 act as a signal probe. In the presence of bPAG9，aptamer 1 and aptamer 2 bind with bPAG9 and forma double aptamer sandwich structure on solid phase carrier. Fe-MIL-101 coupled to aptamer 2 enable to catalyze the colorless peroxidase substrate to yield a colorimetric output signal. Thus，the qualitative and quantitative analysis of bPAG9 could be realized by the change of colorimetric signal. The results showed that under the optimal conditions，the linear detection range were 0.5-50 ng/mL for bPAG9 in buffered solution，with a limit of detection（LOD） of 0.13 ng/mL. The average recoveries of bPAG9 in blank serum samples were in the range of 89.4%-104%，and the relative standard deviations were less than 10%. Subsequently，the serum samples from Holstein cows at 28 days after insemination were analyzed by the developed assay. The test results were confirmed with ultrasonography and compared with PAG-ELISA test kit. The diagnostic sensitivity，specificity，and accuracy of the developed assay were 92.5%，90.0% and 91.5%，respectively.The developed assay has such advantages as simple，noninvasive，and economical. It has a good application prospect in the field of cow early pregnancy detection.

关键词

妊娠相关糖蛋白适配体金属有机框架妊娠诊断奶牛

Keywords

pregnancy associated glycoproteinaptamermetal organic frameworkpregnancy diagnosiscow

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