食品与发酵工业 >

2023 , Vol. 49 >Issue 1: 267 - 272

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.030417

分析与检测

适配体识别-化学发光技术检测福氏志贺菌

  • 陈威风 ,
  • 李晓雪 ,
  • 裴兰梅 ,
  • 常惟丹 ,
  • 崔丽伟 ,
  • 潘春梅
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  • (河南牧业经济学院 食品与生物工程学院,河南 郑州,450046)
第一作者：博士,讲师(潘春梅教授为通信作者,E-mail:pancm7570@163.com)

收稿日期: 2021-12-12

  修回日期: 2022-03-22

  网络出版日期: 2023-02-14

基金资助

河南省科技攻关项目(212102310259);河南省高等学校重点科研项目(22B550003,22B150005)

收起

Detection of Shigella frigneri based on aptamer recognition and chemiluminescence technique

  • CHEN Weifeng ,
  • LI Xiaoxue ,
  • PEI Lanmei ,
  • CHANG Weidan ,
  • CUI Liwei ,
  • PAN Chunmei
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  • (School of Food and Bioengineering,Henan university of animal husbandry and economy, Zhengzhou 450046, China)

Received date: 2021-12-12

  Revised date: 2022-03-22

  Online published: 2023-02-14

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摘要

福氏志贺菌(Shigella flexneri)是一种危害人类健康的食源性致病菌。该实验首先制备鲁米诺功能化花状纳米金和氨基化磁性纳米Fe3O4,然后将福氏志贺菌适配体的互补序列(FSZ-2HH)连接在鲁米诺功能化花状纳米金的表面构建信号探针,并将福氏志贺菌适配体(FSZ-1)通过亲和素的桥接作用修饰到氨基化磁性纳米Fe3O4表面构建捕获探针。当目标物存在时,目标物和信号探针会竞争性结合捕获探针,进而通过测定化学发光来达到检测目的。实验结果表明:在浓度2.6×10~2.6×105 CFU/mL,福氏志贺菌的浓度与化学发光强度呈良好的线性关系,检出限为12 CFU/mL,特异性良好。该实验建立的方法为食品中福氏志贺菌的快速高灵敏检测提供了良好思路。

关键词： 福氏志贺菌; 适配体传感器; 化学发光; 磁分离技术

本文引用格式

陈威风 , 李晓雪 , 裴兰梅 , 常惟丹 , 崔丽伟 , 潘春梅 . 适配体识别-化学发光技术检测福氏志贺菌[J]. 食品与发酵工业, 2023 , 49(1) : 267 -272 . DOI: 10.13995/j.cnki.11-1802/ts.030417

Abstract

Shigella flexneri is a foodborne pathogen highly threatening human health. Firstly, luminol functionalized flower-like gold nanoparticles and amino magnetized nano-Fe3O4 were prepared. Afterward, the complementary sequence of S. flexneri aptamer (FSZ-2HH) was connected to the surface of the luminol functionalized flower-like gold nanoparticles to construct a signal probe whereas the S. flexneri aptamer (FSZ-1) was modified to the amino magnetic nano-Fe3O4 surface by avidin bridging to construct a capture probe. When the Shigella flexneri is present, it will compete with the signal probe for binding the capture probe, whereby the bacteria could be detected via measuring the chemiluminescence. The results showed that in the range of 2.6×101 to 2.6×105 CFU/mL, there was a good linear relationship between the concentration of S. flexneri and the chemiluminescence intensity, with good specificity and a detection limit of 12 CFU/mL. This method provides a potential tool for the rapid and sensitive detection of S. flexneri in foods.

Key words： Shigella flexneri; adapter sensor; chemiluminescence; magnetic separation technology

参考文献

[1] FRITAH S, LHOCINE N, GOLEBIOWSKI F, et al.Sumoylation controls host anti-bacterial response to the gut invasive pathogen Shigella flexneri[J].EMBO Reports, 2014, 15(9):965-972.
[2] ASHIDA H, SUZUKI T, SASAKAWA C.Shigella infection and host cell death:A double-edged sword for the host and pathogen survival[J].Current Opinion in Microbiology, 2021, 59:1-7.
[3] RANALLO R T, KAMINSKI R, BAQAR S, et al.Oral administration of live Shigella vaccine candidates in rhesus monkeys show no evidence of competition for colonization and immunogenicity between different serotypes[J].Vaccine, 2014, 32(15):1 754-1 760.
[4] YANG C J, LI P, ZHANG X J, et al.Molecular characterization and analysis of high-level multidrug-resistance of Shigella flexneri serotype 4 s strains from China[J].Scientific Reports, 2016, 6:29124.
[5] LAVU P S, MONDAL B, RAMLAL S.Selection and characterization of cell surface specific aptamer and development of fluorescence assay for detection of Shigella flexneri from water samples[J].Journal of Fluorescence, 2021, 31(3):685-693.
[6] SHAHIN K, ZHANG L L, DELFAN A S, et al.Effective control of Shigella contamination in different foods using a novel six-phage cocktail[J].LWT, 2021, 144:111137.
[7] SUN X H, XU Q, PAN Y J, et al.A loop-mediated isothermal amplification method for rapid detection of Vibrio parahaemolyticus in seafood[J].Annals of Microbiology, 2012, 62(1):263-271.
[8] ZHU D, YAN Y R, LEI P H, et al.A novel electrochemical sensing strategy for rapid and ultrasensitive detection of Salmonella by rolling circle amplification and DNA-AuNPs probe[J].Analytica Chimica Acta, 2014, 846:44-50.
[9] SHAO Y C, ZHU S M, JIN C C, et al.Development of multiplex loop-mediated isothermal amplification-RFLP (mLAMP-RFLP) to detect Salmonella spp.and Shigella spp.in milk[J].International Journal of Food Microbiology, 2011, 148(2):75-79.
[10] WANG Z Y, YANG Q, ZHANG Y Z, et al.Saltatory rolling circle amplification (SRCA):A novel nucleic acid isothermal amplification technique applied for rapid detection of Shigella spp.in vegetable salad[J].Food Analytical Methods, 2018, 11(2):504-513.
[11] NGUYEN N T, PARK R M, KIM Y H, et al.Detection and discrimination of Shigella sonnei and Shigella flexneri based on vacuolar responses in Saccharomyces cerevisiae[J].Journal of Biotechnology, 2018, 287:1-7.
[12] ZHAO G Y, ZHAN X J, DOU W C.A disposable immunosensor for Shigella flexneri based on multiwalled carbon nanotube/sodium alginate composite electrode[J].Analytical Biochemistry, 2011, 408(1):53-58.
[13] 蒋艳, 余姓鸿, 谢礼, 等.化学发光免疫方法在食品安全检测中的应用及展望[J].食品安全质量检测学报, 2020, 11(20):7 603-7 609.
JIANG Y, YU X H, XIE L, et al.Application and prospect of chemiluminescence immunoassay in food safety detection[J].Journal of Food Safety & Quality, 2020, 11(20):7 603-7 609.
[14] 何华红, 张陆勇, 何玲, 等.化学发光分析及其在药理学上的应用[J].药学进展, 2004, 28(6):257-262.
HE H H, ZHANG L Y, HE L, et al.Chemiluminescence analysis and its application in pharmacology[J].Progress in Pharmaceutical Sciences, 2004, 28(6):257-262.
[15] 马超. 基于纳米磁分离和化学发光的肝炎分子检测新技术研究[D].南京:东南大学, 2018.
MA C.Novel methods for molecule detection of hepatitis viruses based on magnetic separation and chemiluminescence[D].Nanjing:Southeast University, 2018.
[16] 李蕴. 纳米磁分离技术在食品微生物快速检测领域的应用[J].黑龙江科技信息, 2016(31):107.
LI Y.Application of nanomagnetic separation technology in the rapid detection of food microbes[J].Heilongjiang Science and Technology Information, 2016(31):107.
[17] CHU S, LIM B, CHOI C.Application status and prospect of magnetic separation technology for wastewater treatment[J].Journal of Korean Society on Water Environment, 2020, 36(2):153-163.
[18] BINANDEH M, KARIMI F, ROSTAMNIA S.Application of magnetic nanoparticles by comparing the absorbance and stabilization of biomolecules DNA-C, L by the electrophoretic detection[J].International Journal of Health Sciences, 2021, 15(2):3-8.
[19] KARGINA Y V, ZINOVYEV S V, PEREPUKHOV A M, et al.Silicon nanoparticles with iron impurities for multifunctional applications[J].Functional Materials Letters, 2020, 13(4):2040007.
[20] SAVVIDOU M G, FERRARO A, HRISTOFOROU E, et al.Incorporation of magnetic nanoparticles into protoplasts of microalgae Haematococcus pluvialis:A tool for biotechnological applications[J].Molecules (Basel, Switzerland), 2020, 25(21):5068.
[21] 刘莹, 程立, 张博.基于新型磁分离技术的高分辨率DNA片段筛选技术[J].生物技术通报, 2021, 37(10):257-265.
LIU Y, CHENG L, ZHANG B.High-resolution DNA screening based on novel magnetic separation technology[J].Biotechnology Bulletin, 2021, 37(10):257-265.
[22] 于颖, 杨秀娜, 杨振宇, 等.磁分离技术在石油及化工领域的应用[J].石油炼制与化工, 2021, 52(1):20-26.
YU Y, YANG X N, YANG Z Y, et al.Application of magnetic separation technology in petroleum and chemical industry[J].Petroleum Processing and Petrochemicals, 2021, 52(1):20-26.
[23] ZHU W H, LI Z J, LIU X X, et al.Determination of Shigella flexneri by a novel fluorescent aptasensor[J].Analytical Letters, 2015, 48(18):2 870-2 881.
[24] KHOSROSHAHI M E, GHAZANFARI L.Synthesis and functionalization of SiO2 coated Fe3O4 nanoparticles with amine groups based on self-assembly[J].Materials Science and Engineering:C, 2012, 32(5):1 043-1 049.
[25] LI F, CUI H.A label-free electrochemiluminescence aptasensor for thrombin based on novel assembly strategy of oligonucleotide and luminol functionalized gold nanoparticles[J].Biosensors and Bioelectronics, 2013, 39(1):261-267.
[26] LUO J L, WANG J P, MATHEW A S, et al.Ultrasensitive detection of Shigella species in blood and stool[J].Analytical Chemistry, 2016, 88(4):2 010-2 014.
[27] FENG J L, SHEN Q, WU J J, et al.Naked-eyes detection of Shigella flexneri in food samples based on a novel gold nanoparticle-based colorimetric aptasensor[J].Food Control, 2019, 98:333-341.
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