基于卵黄抗体-金属有机框架复合探针的电化学免疫传感器快速检测金黄色葡萄球菌

引用本文: 王家琪, 吴佳怡, 林建原, 李天华, 李登峰, 干宁. 基于卵黄抗体-金属有机框架复合探针的电化学免疫传感器快速检测金黄色葡萄球菌[J]. 分析化学, 2021, 49(2): 197-206. doi: 10.19756/j.issn.0253-3820.201566 shu

Citation: WANG Jia-Qi, WU Jia-Yi, LIN Jian-Yuan, LI Tian-Hua, LI Deng-Feng, GAN Ning. Rapid Detection of Staphylococcus aureus by An Electrochemical Immunosensor Based on Egg Yolk Antibody-Metal Organic Framework Composite Probe[J]. Chinese Journal of Analytical Chemistry, 2021, 49(2): 197-206. doi: 10.19756/j.issn.0253-3820.201566 shu

基于卵黄抗体-金属有机框架复合探针的电化学免疫传感器快速检测金黄色葡萄球菌

王家琪 ^1, ,
吴佳怡 ^1, ,
林建原 ^3, ,
李天华 ^{1,
,} ,
李登峰 ^2, ,
干宁 ^{1,
,}

1.
宁波大学材料科学与化学工程学院, 宁波 315211;
2.
宁波大学海洋学院, 宁波 315211;
3.
浙江万里学院生物与环境分院, 宁波 315200

通讯作者: 李天华,E-mail:litianhua@nbu.edu.cn; 干宁,E-mail:ganning@nbu.edu.cn

基金项目:
国家自然科学基金项目（No.21974074）、浙江省自然科学基金项目（Nos.LY19B050001，LY18B070004，2019A610184）、浙江省基础公益计划研究项目（Nos.KF2019001，LGN18H300001）和宁波大学王宽诚幸福基金资助。

摘要: 金黄色葡萄球菌（S.aureus）污染是影响食品质量安全的重要因素，因此迫切需要开发快速、简便的现场检测食品中S.aureus的分析方法。本研究采用灭活S.aureus免疫无特定病原体（SPF）蛋鸡，从产蛋中纯化出卵黄抗体（IgY），其可以高效、特异性地捕获S.aureus。将IgY固定在丝网印刷电极表面，可用于食品中S.aureus的选择性分离富集。在UiO-66金属有机框架（MOF）材料表面共价连接二茂铁（Fc）和苯硼酸（BPA），获得了MOF@Fc-BPA复合探针。此探针与电极上的IgY共同识别并捕获S.aureus，并在电极上产生Fc的氧化还原信号。在最佳反应条件下，电化学信号强度与S.aureus浓度的对数值成正比，检测范围为10~10⁹ cfu/mL，检出限为3 cfu/mL，检测时间为20 min。采用此传感器检测了养殖虾体内的S.aureus，检测结果与标准ELISA方法的结果一致，但检测时间明显缩短，检出限比ELISA方法低两个数量级。此传感器具有良好的现场快速筛查S.aureus的能力。

关键词:

English

Rapid Detection of Staphylococcus aureus by An Electrochemical Immunosensor Based on Egg Yolk Antibody-Metal Organic Framework Composite Probe

1.
School of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, China;
2.
College of Oceanography, Ningbo University, Ningbo 315211, China;
3.
College of Biology and Environment, Zhejiang Wanli University, Ningbo 315200, China

Corresponding author: LI Tian-Hua, ; GAN Ning,

Received Date: 20 September 2020
Revised Date: 25 November 2020
Fund Project:
Supported by the National Natural Science Foundation of China (No.21974074), the Zhejiang Provincial Natural Science Foundation of China (Nos.LY19B050001, LY18B070004, 2019A610184), the Zhejiang Basic Public Welfare Program Research Project (Nos.KF2019001, LGN18H300001) and K C Wong Magna Fund in Ningbo University.

Abstract: Staphylococcus aureus (S.aureus) contamination is an important factor affecting food quality and safety. Therefore, it is urgent to develop a quick and easy on-site analysis method for detecting S.aureus in food. In this work, inactivated S.aureus was used to immunize specific pathogen free (SPF) hens, and the yolk antibody (IgY) was purified from the eggs wich could efficiently and specifically capture S.aureus. IgY modified on the surface of the screen-printed electrode could be used for the selective separation and enrichment of S.aureus in the sample. Ferrocene (Fc) and phenylboronic acid (BPA) were covalently connected on the surface of UiO-66 nano-metal organic framework material (MOF) to obtain a MOF@Fc-BPA composite probe. The prepared probe and the IgY on the electrode jointly identified and captured S.aureus, and then generated a redox signal of Fc on the electrode. Under the optimal reaction conditions, the electrochemical signal intensity was proportional to the logarithm of the S.aureus concentration, the detection range was 10-10⁹ cfu/mL, the detection limit was 3 cfu/mL, and the detection time was 20 min only. In addition, we used this sensor to detect S.aureus in cultured shrimp, and the results obtained were consistent with that of standard ELISA, while the detection time was greatly shortened, and the detection limit was two orders of magnitude lower than the ELISA method. The results indicated that the immunosensor was able to quickly screen S.aureus on site.

Key words:

Egg yolk antibody
/ Staphylococcus aureus
/ Ferrocene/phenylboronic acid-metal organic framework composite probe
/ Electrochemical immunosensor
/ On-site rapid detection

1. [1]
  VELUSAMY V, ARSHAK K, KOROSTYNSKA O, OLIWA K, ADLEY C. Biotechnol. Adv., 2010, 28(2):232-254.
2. [2]
  NGUYEN A T, TALLENT S M. J. AOAC Int., 2018, 101(4):1127-1134.
3. [3]
  KADARIYA J, SMITH T C, THAPALIYA D. Biomed. Res. Int., 2014, 2014:827965.
4. [4]
  GB 4789.10-2016, National Food Safety Standard Food Microbiology Inspection Staphylococcus Aureus Inspection. National Standards of the People's Republic of China. 食品安全国家标准食品微生物学检验金黄色葡萄球菌检验. 中华人民共和国国家标准. GB 4789.10-2016.
5. [5]
  LING Y, ZHU Y, FAN H, ZHA H, YANG M, WU L, CHEN H, LI W, WU Y, CHEN H. J. Biomed. Nanotechnol., 2019, 15(6):1290-1298.
6. [6]
  NEMR C R, SMITH S J, LIU W, MEPHAM A H, MOHAMADI R M, LABIB M, KELLEY S O. Anal. Chem., 2019, 91(4):2847-2853.
7. [7]
  SHAN Y, XU C, WANG M, ZHU Z, WU F G, SHI Z, CUI Q, ARUMUGAM G M. Talanta, 2019, 192:154-159.
8. [8]
  AOYAMA S, MONDEN K, AKIYAMA Y, YAMADA M, SEKI M. Anal. Chem., 2019, 91(21):13377-13382.
9. [9]
  GU Ye-Hua, LI Deng-Feng, ZHOU Yong-Qiang. J. Ningbo Univ. (Sci. Technol. Ed.), 2018, 5(31):1-6. 顾叶华, 李登峰, 周永强. 宁波大学学报(理工版), 2018, 5(31):1-6.
10. [10]
  THIRUMALAI D, AMBI S V, VIEIRA-PIRES R S, ZHENG X Y, SEKARAN S, KRISHNAN U. Int. J. Biol. Macromol., 2019, 136:755-763.
11. [11]
  LI Min-Hui, ZOU Qiang, YANG Shu-Xia, HE Jian-Rong, LIU Yang, HUANG Ya-Lin, CHANG Shan. Chin.J. Biol., 2009, 26(6):80-82. 李敏惠, 邹强, 杨淑霞, 何建荣, 刘阳, 黄雅琳, 常山. 生物学杂志, 2009, 26(6):80-82.
12. [12]
  XIA Xiao-Lei, LI Deng-Feng, PENG Jiao, ZHOU Yong-Qiang, GUO An-Nan, LIU Lian-Guo, CHEN Jiong. J. Ningbo Univ. (Sci. Technol. Ed.), 2016, 29(4):13-19. 夏小磊, 李登峰, 彭姣, 周永强, 郭安南, 刘联国, 陈炯. 宁波大学学报(理工版), 2016, 29(4):13-19.
13. [13]
  YAN Mao-Cang, WANG Yao-Hua, HU Li-Hua, ZHANG Jiong-Ming, LIU Ya-Nan, TANG Ming, LUO Kui, HUANG Xian-Ke, ZHOU Qian-Jin, CHEN Jiong. Chin. Oceans Lakes, 2019, 50(2):443-448. 闫茂仓, 王瑶华, 胡利华, 张炯明, 刘亚南, 唐明, 罗奎, 黄贤克, 周前进, 陈炯. 海洋与湖沼, 2019, 50(2):443-448.
14. [14]
  KIM D K, JANG I K, SEO H C, SHIN S O, YANG S Y, KIM J W. Aquaculture, 2004, 237(1-4):21-30.
15. [15]
  WU C J, WANG H, CHAN Y L, LI T L. Fish Shellfish Immunol., 2011, 30(4-5):1042-1048.
16. [16]
  HUANG S, GAN N, ZHANG X, WU Y, SHAO Y, JIANG Z, WANG Q. Biosens. Bioelectron, 2019, 128:113-121.
17. [17]
  LI Zhu-Heng, GAO Jing-Qing, JIAN Ming-Hong, WANG Zhen-Xin. Chin. J. Anal. Chem., 2018, 46(12):1904-1912. 李铸衡, 高晶清, 简明红, 王振新. 分析化学, 2018, 46(12):1904-1912.
18. [18]
  ZHOU H C, KITAGAWA S. Chem. Soc. Rev., 2014, 43(16):5415-5418.
19. [19]
  ROSTOVTSEV V V, GREEN L G, FOKIN V V, SHARPLESS K B. Angew. Chem. Int. Ed., 2002, 41(14):2596-2599.
20. [20]
  LI P, GE B, OU L M, YAO Z, YU H Z. Sensors (Basel), 2015, 15(8):20543-20556.
21. [21]
  LATOUR R A. J. Biomed. Mater. Res. A, 2015, 103(3):949-958.
22. [22]
  ABBASPOUR A, NOROUZ S F, NOORI A, SOLTANI N. Biosens. Bioelectron., 2015, 68:149-155.
23. [23]
  DUAN N, WU S, CHEN X, HUANG Y, WANG Z. J. Agric. Food Chem., 2012, 60(16):4034-4038.
24. [24]
  GWENAËLLE L B, ISMAÏL F, CHRISTOPHE L. J. Microbiol. Methods, 2004, 59(2):211-221.
25. [25]
  KHATEB H, KLÖS G, MEYER R L, SUTHERLAND D S. ACS Appl. Bio. Mater., 2020, 3(5):3066-3077.

扫一扫看文章

计量

PDF下载量: 19
文章访问数: 737
HTML全文浏览量: 96

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

基于卵黄抗体-金属有机框架复合探针的电化学免疫传感器快速检测金黄色葡萄球菌

通讯作者: 李天华,E-mail:litianhua@nbu.edu.cn; 干宁,E-mail:ganning@nbu.edu.cn

English

Rapid Detection of Staphylococcus aureus by An Electrochemical Immunosensor Based on Egg Yolk Antibody-Metal Organic Framework Composite Probe

Corresponding author: LI Tian-Hua, ; GAN Ning,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于卵黄抗体-金属有机框架复合探针的电化学免疫传感器快速检测金黄色葡萄球菌

通讯作者: 李天华,E-mail:litianhua@nbu.edu.cn; 干宁,E-mail:ganning@nbu.edu.cn

English

Rapid Detection of Staphylococcus aureus by An Electrochemical Immunosensor Based on Egg Yolk Antibody-Metal Organic Framework Composite Probe

Corresponding author: LI Tian-Hua, ; GAN Ning,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs