食品安全問題是各國政府和科學(xué)家面臨的主要挑戰(zhàn)之一。真菌毒素是由真菌產(chǎn)生的有毒次生代謝產(chǎn)物,能夠?qū)е氯祟惡蛣游锼劳�。目�?人們已從農(nóng)業(yè)科學(xué),環(huán)境科學(xué)、免疫學(xué)、藥理學(xué)和醫(yī)學(xué)等多方面對真菌毒素展開了認(rèn)真的研究。由于色譜法和傳統(tǒng)免疫分析法的不足,所以需要開發(fā)創(chuàng)新和穩(wěn)定的方法來對真菌毒素進(jìn)行適宜、靈敏和快速的檢測。正因如此,各國都在研制和發(fā)展一次性適配體傳感器/生物傳感器,用以識別真菌毒素。近年來,研究者們開發(fā)了多種針對各類真菌毒素的熒光適配體傳感器,而時間分辨熒光技術(shù)與適配體相結(jié)合的應(yīng)用才剛剛出現(xiàn)。時間分辨熒光技術(shù)（TRF）可以有效消除背景熒光,具有較高的噪比。通過設(shè)置一定的延遲時間,可以區(qū)分樣品中所含熒光物質(zhì)的熒光壽命,從而提高檢測的靈敏度�；诖�,本研究中,合成了幾種時間分辨的熒光納米材料并對其進(jìn)行表征,并結(jié)合多種納米材料（如磁性納米材料、二硫化鎢、石墨烯氮化碳納米片）及相應(yīng)技術(shù)（時間分辨熒光技術(shù)、磁分離技術(shù)、滾環(huán)擴(kuò)增技術(shù)）等,構(gòu)建了一系列快速,簡單,靈敏,高效的適配體傳感器用于檢測多種真菌毒素。首先,基于已報道的玉米赤霉烯酮（ZEN）適配體序列,構(gòu)建了一種新穎、高靈敏、強(qiáng)親和性的時間分辨... 

【文章來源】：江南大學(xué)江蘇省 211工程院校 教育部直屬院校

【文章頁數(shù)】：148 頁

【學(xué)位級別】：博士

【文章目錄】：
ACKNOWLEDGEMENTS
DEDICATION
LIST OF ABBREVIATIONS
Abstract
摘要
Chapter 1 General introduction and review of literature
    1.1 Mycotoxins: General information
        1.1.1 Zearalenone (ZEN)
        1.1.2 Fumonisin
        1.1.3 Ochratoxin A
        1.1.4 Trichothecenes A (T-2 toxin)
        1.1.5 Aflatoxin
    1.2 Natural co-occurrence of mycotoxins
    1.3 Conventional analytical techniques for mycotoxin detection
        1.3.1 Chromatographic techniques
        1.3.2 Immunological methods
    1.4 Aptasensors for mycotoxins detection
        1.4.1 Elements of aptasensors
            1.4.1.1 Aptamers: General information and aptamers selected for mycotoxins
            1.4.1.2 Nanomaterials
                1.4.1.2.1 Quantum dots (QDs)
                1.4.1.2.2 Magnetic nanoparticles
                1.4.1.2.3 Silica nanoparticles (Si-NPs)
                1.4.1.2.4 Carbon nanomaterial
                1.4.1.2.5 Metal nanomaterial
                1.4.1.2.6 Up conversion nanoparticles (UCNPs)
    1.5 Nanoparticles based aptasensors for mycotoxins detection
        1.5.1 Colorimetric sensors
        1.5.2 Electrochemical sensors
        1.5.3 Fluorometric sensors
    1.6 Time-resolved fluorescence technique (TRFL)
    1.7 Time-gated luminescence technique (TGL)
    1.8 Principle of time-resolved fluorescence technique
    1.9 Ln~(3+)-doped fluorescent inorganic nanoparticles for time-resolved biosensing
    1.10 Ln~(3+)-doped silica nanoparticles for time-resolved biosensing
    1.11 Other techniques used in this research
        1.11.1 Magnetic separation technique
        1.11.2 Rolling circle amplification (RCA)
            1.11.2.1 Rolling circle amplification process
            1.11.2.2 Advantages of RCA
            1.11.2.3 RCA-based fluorescence detection assay
            1.11.2.4 RCA based fluorescence detection assay for mycotoxin
    1.12 Other 2D nanomaterials used in this research
        1.12.1 Tungsten disulfide nanomaterial (WS2)
        1.12.2 Graphitic carbon nitride (g-C_3N_4)
    1.13 Importance of research topic
    1.14 Main objectives of the research
    1.15 Technical route of the research
    1.16 References
CHAPTER 2: A novel bioassay based on aptamer-functionalized magnetic nanoparticle for the detection of zearalenone using time resolved-fluorescence Na YF_4: Ce/Tb nanoparticles as signalprobe
    2.1 Introduction
    2.2 Materials and methods
        2.2.1 Materials and apparatus
        2.2.2 Preparation of amine-functionalized Fe3O4 magnetic nanoparticles
        2.2.3 Preparation of Na YF_4: Ce/Tb nanoparticles
        2.2.4 Synthesis of signal probe assembly and ZEN measurement
        2.2.5 Sample preparation and measurement
    2.3 Results and discussion
        2.3.1 Principle-based on TRFL-NPs and MNPs for detection of ZEN
        2.3.2 Characterization of the prepared Na YF_4: Ce/Tb and amine-functionalized magneticnanoparticles
        2.3.3 Characteristics of avidin-conjugated TRFL-NPs and MNPs
        2.3.4 Optimization for assay
            2.3.4.1 Aptamers concentration (n M)
            2.3.4.2 Volume of apt-MNPs (μL)
        2.3.5 Analytical performance
        2.3.6 Specificity assay
        2.3.7 Analytical applications
    2.4 Conclusions
    2.5 References
CHAPTER 3: Simultaneous Detection of Fumonisin B1 and Ochratoxin A using Dual-Colour Time Resolved Luminescent Nanoparticles (Na YF_4: Ce, Tb and NH_2-Eu/DPA@Si O_2) asLabels
    3.1 Introduction
    3.2 Materials and methods
        3.2.1 Reagents
        3.2.2 Apparatus
        3.2.3 Synthesis of amine-functionalized Fe3O4 MNPs
        3.2.4 Synthesis of Na YF_4: Ce, Tb NPs
        3.2.5 Synthesis of NH2-Eu/DPA@Si O2 NPs
        3.2.6 Synthesis of signal probe (MNPs-Apt_1/Na YF_4: Ce, Tb-c DNA1 and MNPs-Apt_2/NH_2-Eu/DPA@Si O_2-c DNA_2)
        3.2.7 Procedure for simultaneous detection
        3.2.8 Preparation and measurement of real sample
    3.3 Results and discussion
        3.3.1 Detection principle
        3.3.2 Characterizations of TRF-NPs and MNPs
        3.3.3 Characterizations of NPs conjugated to avidin and aptamer
        3.3.4 Optimization of experimental conditions
            3.3.4.1. Effect of aptamers concentrations
            3.3.4.2. Effect of volume of MNPs-aptamer conjugates
            3.3.4.3. Effect of incubation time
        3.3.5 Analytical performance
        3.3.6 Specificity evaluation
        3.3.7 Analytical applications
    3.4 Conclusion
    3.5 References
CHAPTER 4: A“turn on”aptasensor for simultaneous detection of zearalenone, trichothecenes Aand aflatoxin B1 mycotoxins using WS2 as a quencher for time-resolved fluorescence
    4.1 Introduction
    4.2 Materials and methods
        4.2.1 Reagents
        4.2.2 Instrumentation
        4.2.3 Preparation of amine functionalized Ln~(3+) doped KYF_4 NPs
        4.2.4 Preparation of aptamer modified KYF_4: Ln~(3+) multicolor bioprobe
        4.2.5 Procedure for simultaneous detection of mycotoxins
        4.2.6 Sample preparation
    4.3 Results and discussion
        4.3.1 Detection scheme
        4.3.2 Characterization of Ln~(3+) doped KYF_4 NPs and WS2
        4.3.3 Characterization of NPs conjugated to avidin and aptamers
        4.3.4 Optimization of developed aptasensor
        4.3.5 Analytical performance
        4.3.6 Selectivity of assay
        4.3.7 Real food sample analysis
    4.4 Conclusion
    4.5 References
CHAPTER 5: Fluorescence detection of aflatoxin M_1 using rolling circle amplification and carbonnitride nanosheet as quencher
    5.1 Introduction
    5.2 Materials and methods
        5.2.1 Materials and reagents
        5.2.2 Instrumentation
        5.2.3 Synthesis of KYF_4: Eu~(+3) NPs
        5.2.4 Synthesis of signal probe (KYF_4: Eu~(+3)-c DNA)
        5.2.5 Preparation of ultrathin g-C_3N_4 nanosheets
        5.2.6 Procedure for circularization of rolling circle template (RCT) DNA
        5.2.7 Procedure for rolling circle amplification (RCA) reaction
        5.2.8 Procedure for fluorescence measurement of RCA product hybridized with signal probeand g-C_3N_4
        5.2.9 Analytical procedure
        5.2.10 Sample preparation
    5.3. Result and discussion
        5.3.1. Sensing principle
        5.3.2. Characterization of KYF_4: Eu~(+3) NPs and ultrathin g-C_3N_4 nanosheet
        5.3.3 Characteristics of signal probe (TRFNPs-c DNA) avidin and c DNA
        5.3.4 Characterization of circularization of rolling circle template (RCT) DNA
        5.3.5 Characterization of rolling circle amplification (RCA) product
        5.3.6 Optimization of experimental conditions
        5.3.7 Analytical performance
        5.3.8 Specificity and selectivity
        5.3.9 Analytical application in milk sample
    5.4 Conclusion
    5.5 References
Chapter 6: General conclusion and recommendation
    General conclusion
    Key innovations
    Recommendations
    List of Publications


【參考文獻(xiàn)】：
期刊論文
[1]玉米赤霉烯酮單克隆抗體的制備及間接競爭ELISA檢測方法的建立[J]. 王元凱,王君,王雨晨,陳志飛,嚴(yán)亞賢,郝倩雯,李樹清,于翠,楊翠云,孫建和.  微生物學(xué)通報. 2011(12)
[2]應(yīng)用生物素-鏈霉親和素的酶聯(lián)免疫吸附法檢測谷物中的玉米赤霉烯酮[J]. 馬智鴻,黃飚,屠薔,高蕾,張玨,王柯.  安徽農(nóng)業(yè)科學(xué). 2009(15)



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