CN112881352B - 用于沙门氏菌检测和消杀的核酸适配体-量子点生物传感器、其制备方法及应用 - Google Patents
用于沙门氏菌检测和消杀的核酸适配体-量子点生物传感器、其制备方法及应用 Download PDFInfo
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Abstract
本发明公开了一种用于沙门氏菌检测和消杀的核酸适配体‑量子点生物传感器、其制备方法及应用，属于食品致病菌的检测及灭活技术领域。本发明的生物传感器是在沉积有聚多巴胺(PDA)涂层的Fe3O4纳米粒子表面连接经量子点标记的沙门氏菌核酸适配体Aptamer C；并将修饰有荧光素的互补链Strand A与上述Aptamer C配对；用所述连接有Aptamer C和Strand A的Fe3O4纳米粒子进行沙门氏菌的检测。本发明的生物传感器的检测灵敏度为10CFU/100g，检测范围为10‑104CFU/100g，具有很好的灵敏度和特异性；灭菌程度达95％以上，效果显著。
Description
技术领域
本发明属于食品致病菌的检测及灭活技术领域，具体涉及一种用于沙门氏菌检测和消杀的核酸适配体-量子点生物传感器、其制备方法及应用。
背景技术
食品致病菌是可以引起食物中毒或以食品为传播媒介的致病性细菌。致病性细菌直接或间接污染食品及水源，人经口感染可导致肠道传染病、食物中毒以及畜禽传染病的流行。因此食源性致病菌是导致食品安全问题的重要来源。
食源性致病菌对人体健康的影响：(1)引起急性中毒：在一般情况下，食源性致病菌常引起急性中毒，轻者多以急性肠胃炎症状出现，如呕吐、恶心、腹痛、腹泻、发烧等，经过治疗可以恢复健康；但重者可出现呼吸、循环、神经等系统症状，抢救及时可转危为安，若耽误时机可能危及生命。有的急性中毒，虽经千方百计治疗，但仍给中毒者留下后遗症。(2)慢性中毒或潜在性危害：有些变质食物中的有毒物质含量少，或者由于本身毒性作用的特点，并不引起急性中毒，但长期使用，往往可造成慢性中毒，甚至可以表现出致癌、致畸、致突变的作用。使用腐败变质、霉变食物除了可以引起急性中毒外，还具有极其严重的潜在危害。
沙门氏菌是我国食源性疾病中引起食物中毒的最常见致病菌，约占中毒病例的70％-80％，而以肉、蛋、奶等畜产品被沙门氏菌污染所引起的中毒可占90％以上。沙门氏菌感染症为人畜共患感染性疾病，主要由食用遭受污染的食物导致，摄入含毒食品后，症状一般在12～14h内出现，有些潜伏期较长，典型症状包括发热、恶心、呕吐、腹泻及腹部绞痛，还伴有乏力、肌肉酸痛、视觉模糊、躁动不安和嗜睡等，但一般在发热后72h内会好转。沙门氏菌还可能会引起肠伤寒，肠胃炎和败血症疾病，而婴儿、老年人、免疫功能低下的患者则可能因沙门氏菌进入血液而出现严重且危及生命的菌血症，少数还会合并脑膜炎或骨髓炎。
沙门氏菌是美国食物中毒致死的主要原因。美国人对这种病菌一点都不陌生，每年全国大约报告40000例沙门氏菌感染病例。但实际的感染人数可能要达20倍以上，因为许多轻型病人可能未确诊，据不完全统计，每年大约有1000人死于急性沙门氏菌感染。致病菌的感染不但会造成巨大的经济损失，而且严重威胁人民群众的健康。因此，食品的致病菌检测和灭菌技术越来越重要。检测沙门氏菌一直以传统检测方法为主，非选择性和选择性增菌、可疑细菌分离等常规方法虽然经典可靠，但程序复杂、十分繁琐，不仅费时费力而且敏感性和特异性较差，漏检率较高。而免疫荧光法、酶联免疫吸附试验(ELISA)、聚合酶链反应(PCR)技术等方法需要使用指定的仪器设备、具备相应的试验条件和技能，难以在基层推广。因此迫切需要在此领域开展深入的研究工作，建立快速、有效的方法检测食品中的沙门氏菌并将其杀灭。
发明内容
针对现有技术中存在的问题，本发明的目的在于提供一种用于沙门氏菌检测和消杀的核酸适配体-量子点生物传感器、其制备方法及应用；本发明的核酸适配体-量子点生物传感器是在沉积PDA的Fe3O4纳米粒子表面连接致病菌的核酸适配体Aptamer C链(链上带碳量子点)，并将修饰有荧光素的互补链Strand A与上述Aptamer C链配对，通过开关荧光进行致病菌检测；在量子点显示蓝绿色荧光区域进行近红外照射，借助Fe3O4纳米粒子表面PDA涂层增强的光热效应，可杀死沙门氏菌。本发明的上述生物传感器及杀菌方法可以解决现有检测及杀灭沙门氏菌技术中存在的灵敏度低、检测方法繁琐、仪器设备要求高、杀菌不彻底等问题。
为了达到上述目的，本发明采用如下技术方案：
用于沙门氏菌检测和消杀的核酸适配体-量子点生物传感器，是在沉积有PDA涂层的Fe3O4纳米粒子表面连接经量子点标记的沙门氏菌核酸适配体Aptamer C；并将修饰有荧光素的互补链Strand A与上述Aptamer C配对；
所述沙门氏菌核酸适配体Aptamer C的核苷酸序列如SEQ ID NO.1所示；
所述互补链Strand A的核苷酸序列如SEQ ID NO.2所示。
上述用于沙门氏菌检测和消杀的核酸适配体-量子点生物传感器的制备方法，步骤如下：
(1)将经量子点标记的Aptamer C链活化并与沉积有PDA涂层的Fe3O4纳米粒子连接：取160μL 2μM的量子点标记的Aptamer C链与30μL 10mg/L的NHS和30μL 10mg/L的EDC混合进行活化；将160μL 2μM活化的量子点标记的Aptamer C链与沉积有PDA涂层的Fe3O4纳米粒子以体积比为3∶1的比例混合；在室温下温育30分钟并轻轻摇动后，以10,000rpm离心25分钟除去游离的链，并用pH 7.0的10mM PBS冲洗3次，将获得的结合物重新分散在500μLPBS缓冲液中；
(2)将修饰有荧光素的互补链Strand A通过碱基互补配对作用借助氢键与步骤(1)获得的结合物的Aptamer C链进行结合：将200μL 2μM的Strand A加入步骤(1)结合物分散的500μL PBS缓冲液中混合；在室温下温育30分钟并轻轻摇动后，以10,000rpm离心25分钟除去游离的Strand A，并用pH 7.0的10mM PBS冲洗3次，即得。
所述荧光素为FAM或者FITC等可以猝灭量子点的荧光素中的任意一种；
在上述方案的基础上，所述的量子点为碳量子点、石墨烯量子点、核壳类和复合材料的无毒荧光量子点如石墨氮化碳量子点、硼掺杂碳点等中的任意一种，优选的为碳量子点；所述碳量子点是以抗坏血酸为碳源，水热法一步制备粒子尺寸为2-3nm，荧光性能良好的碳量子点。
本发明采用碳点合成的量子点与其他材料合成的量子点相比都更加安全和无毒，且碳量子点具有良好的生物相容性，出色的荧光性(包括光稳定性，耐光漂白性和不闪烁性)，成本效益低，制造方法简单、在水中的溶解度高等优势。另外，还可以采用石墨烯量子点和核壳类以及复合材料类的无毒的荧光量子点进行检测沙门氏菌，这些量子点的优势都是无毒、安全的。
在上述方案的基础上，所述在Aptamer C链上标记量子点的方法为：
将10μM碳量子点与1.6μM硫醇修饰的沙门氏菌适配体Aptamer C链在1mL磷酸盐缓冲溶液(pH 7.4，0.01M)中混合；搅拌12h后，收集制备的碳量子点-适体-缀合物共轭物，并通过以6000rpm离心15min用50％乙醇洗涤3次；即得经量子点标记的Aptamer C链；将上述获得的经量子点标记的Aptamer C链悬浮在0.01M磷酸盐缓冲溶液(pH 7.4)中，以待后续制备沙门氏菌的检测试剂。
在上述方案的基础上，所述Fe3O4纳米粒子由以下方法合成：将4.8g六水合氯化铁和2g四水合氯化亚铁溶于60mL的H2O中，然后加到90mL的氢氧化钠溶液中，搅拌30min之后进行磁分离、水洗得到黑色固体。将80mL(1∶1)的乙醇水溶液、5mL浓氨水和1.5g黑色固体，缓慢加入到14mL正硅酸乙酯的乙醇溶液，氮气保护下，50℃下搅拌反应4h，蒸发乙醇。反应继续进行4h后，将产物水洗至中性后用HCl浸泡两天，在50℃下真空干燥箱中干燥至恒重，得到Fe3O4纳米粒子。
本发明选用Fe3O4合成纳米粒子原因：Fe3O4纳米粒子是一种最常见的磁性纳米材料，毒性低、化学稳定性好、磁饱和强度大、粒径分布范围广，且易于表面修饰。
在上述方案的基础上，在Fe3O4纳米粒子上沉积PDA的方法为：
将Fe3O4纳米粒子(12mg)和多巴胺盐酸盐粉末(7.5mg)混合在30mL Tris-HCl缓冲液(pH 8.5)中进行。Fe3O4纳米粒子沉积1h，2h，3h后的PDA涂层厚度差别不大，因此沉积1h为最佳时间。将混合物置于Eppendorf 5436恒温振荡器中，使其在室温下缓慢但连续摇动。沉积PDA涂层后的磁珠通过磁引力分离，并用去离子水清洗干净(三次)，即得沉积有PDA涂层的Fe3O4纳米粒子。
上述方法制备的核酸适配体-量子点生物传感器在食品中沙门氏菌检测和消杀中的应用。
在上述方案的基础上，所述食品为固相食品；优选的为肉制品、果蔬制品中的一种。
使用上述核酸适配体-量子点生物传感器检测和消杀食品中沙门氏菌的方法，步骤如下：
将上述核酸适配体-量子点生物传感器分散在磷酸盐缓冲盐水中，制备成浓度为0.32μM的溶液，将上述分散液添加到食品表面用于检测其中的沙门氏菌，沙门氏菌因高亲和力作用与Aptamer C链特异性结合，Strand A被菌竞争下来，Aptamer C链上的碳量子点会因失去A链上荧光素的猝灭作用而显示出荧光，通过检测量子点的荧光用于测定沙门氏菌的含量；
在显示荧光的区域，利用1.5W/cm2 808nm近红外光进行持续照射；杀菌完成后将食品置于磁场中或者在磁铁的作用下分离出Fe3O4纳米粒子，并将食品表面修饰有荧光素的Strand A用水洗掉即可。
本发明技术方案的优点：
本发明将修饰有碳量子点的沙门氏菌适配体Aptamer C链及其修饰有荧光素的互补链A链修饰在含有PDA涂层的Fe3O4纳米粒子上，通过开关荧光进行特异性识别和检测沙门氏菌，然后利用PDA增强的光热效应来杀灭沙门氏菌。
本发明方法具有特异性，利用适配体识别沙门氏菌具有灵敏度高、亲和力强的特点；操作简便，无需对待测样本进行复杂的前处理；杀菌效果显著。
特别的，当本发明用于食品中沙门氏菌检测时，所用材料均无毒无害，既能杀灭食源性致病菌，又不会给食品造成不良影响，有利于提高食品品质。
本发明适用于固相食品中沙门氏菌的检测和消杀，比如肉制品、果蔬制品等等，使用范围非常广泛，具有很好的实用性。
本发明借助适配体链和量子点的检测灵敏度为10CFU/100g，检测范围为10-104CFU/100g，具有很好的灵敏度和特异性。且灭菌程度达95％以上，效果显著。
附图说明
图1是本发明方法的检测及杀菌原理示意图；
图2是沉积时间对Fe3O4纳米粒子表面沉积PDA涂层厚度的影响；
图3是不同菌株悬浮液吸光度的检测结果图；
图4是含不同浓度沙门氏菌的肉制品检测示意图；
图5是不同浓度沙门氏菌的检测标准曲线图；
图6是Fe3O4纳米粒子沉积PDA前后进行近红外杀菌的荧光强度对比图。
具体实施方式
在本发明中所使用的术语，除非有另外说明，一般具有本领域普通技术人员通常理解的含义。
下面结合具体实施例，并参照数据进一步详细的描述本发明。以下实施例只是为了举例说明本发明，而非以任何方式限制本发明的范围。
本发明用于沙门氏菌检测和消杀的核酸适配体-量子点生物传感器的工作原理：
参照图1，首先合成Fe3O4纳米粒子，然后将其进行PDA沉积，通过沉积的PDA涂层可增强纳米粒子的光热效应，提高杀菌温度。将合成的Aptamer C链修饰上碳量子点(显示蓝绿色荧光)，通过与PDA的氨基羧基脱水缩合作用可将其连接在Fe3O4纳米粒子上，然后将修饰有荧光素的A链与上述Aptamer C链进行互补配对。由于荧光素的荧光猝灭作用，此时Aptamer C链上的碳量子点不显示蓝绿色荧光。将最终修饰好的Fe3O4纳米粒子用于食品中沙门氏菌的识别检测，由于高亲和力Aptamer C会与沙门氏菌特异性结合，A链被菌竞争下来，Aptamer C链上的碳量子点会失去荧光素的荧光猝灭作用显示蓝绿色荧光。在显示荧光区域进行近红外照射，借助Fe3O4纳米粒子表面PDA涂层增强的光热效应，可杀死沙门氏菌。
实施例1
用于沙门氏菌检测和消杀的核酸适配体-量子点生物传感器，是在沉积有PDA涂层的Fe3O4纳米粒子表面连接经碳量子点标记的沙门氏菌核酸适配体Aptamer C(SEQ IDNO.1)；并将修饰有荧光素FAM的互补链Strand A(SEQ ID NO.2)与上述Aptamer C配对；
所述沙门氏菌核酸适配体Aptamer C的5'修饰有羧基，3'修饰有－SH，具体如下所示：
5'－COOH-AGCAGCACAG-GCGATCCAAGCTTCTTCA－SH-3'
所述修饰有荧光素FAM的互补链Strand A如下所示：
3'－CGCTAGGTTCGAAGAAGT－FAM-5'
所述碳量子点是以抗坏血酸为碳源，水热法一步制备粒子尺寸为2-3nm，荧光性能良好的碳量子点。
上述用于沙门氏菌检测和消杀的核酸适配体-量子点生物传感器的制备方法，步骤如下：
(1)在Aptamer C链上标记量子点：将10μM碳量子点与16μM硫醇修饰的沙门氏菌适配体Aptamer C链(购买自生工生物工程(上海)股份有限公司)在1mL磷酸盐缓冲溶液(pH7.4，0.01M)中混合；搅拌12h后，收集制备的碳量子点-适体-缀合物共轭物，并通过以6000rpm离心15min用50％乙醇洗涤3次；即得经量子点标记的Aptamer C链；将上述获得的经量子点标记的Aptamer C链悬浮在0.01M磷酸盐缓冲溶液(pH 7.4)中，以待后续制备沙门氏菌的检测和消杀试剂。
(2)修饰荧光素FAM的Strand A购买自生工生物工程(上海)股份有限公司。
(3)合成Fe3O4纳米粒子：将4.8g六水合氯化铁和2g四水合氯化亚铁溶于60mL的H2O中，然后加到90mL的氢氧化钠溶液中，搅拌30min之后进行磁分离、水洗得到黑色固体。将80mL(1∶1)的乙醇水溶液、5mL浓氨水和1.5g黑色固体，缓慢加入到14mL正硅酸乙酯的乙醇溶液，氮气保护下，50℃下搅拌反应4h，蒸发乙醇。反应继续进行4h后，将产物水洗至中性后用HCl浸泡两天，在50℃下真空干燥箱中干燥至恒重，得到Fe3O4纳米粒子。
(4)在Fe3O4纳米粒子上沉积PDA：将Fe3O4纳米粒子(12mg)和多巴胺盐酸盐粉末(7.5mg)混合在30mL Tris-HCl缓冲液(pH 8.5)中进行反应。如图2所示，Fe3O4纳米粒子沉积1h，2h，3h后的PDA涂层厚度差别不大，均在70nm左右，因此沉积1h为最佳时间。将混合物置于Eppendorf 5436恒温振荡器中，使其在室温下缓慢但连续摇动。沉积PDA涂层后的磁珠通过磁引力分离，并用去离子水清洗干净(三次)，即得沉积有PDA涂层的Fe3O4纳米粒子。
(5)将经量子点标记的Aptamer C链活化并与沉积有PDA涂层的Fe3O4纳米粒子连接：取160μL 2μM的量子点标记的Aptamer C链与30μL 10mg/L的NHS和30μL 10mg/L的EDC混合进行活化。将上述活化的量子点标记的Aptamer C链与沉积有PDA涂层的Fe3O4纳米粒子以体积比为3∶1的比例混合；在室温下温育30分钟并轻轻摇动后，以10,000rpm离心25分钟除去游离的链，并用pH 7.0的10mM PBS冲洗3次，将获得的结合物重新分散在500μL PBS缓冲液中，温度为4℃。通过Fe3O4纳米粒子和适体的浓度来估计在每个Fe3O4纳米粒子上修饰的适体的数目。根据修饰后260nm处吸光度的降低来确定Fe3O4纳米粒子上修饰的适体的浓度。
(6)将修饰有荧光素FAM的互补链Strand A通过碱基互补配对作用借助氢键与上述Aptamer C链进行结合，所述Strand A浓度为2μM，用量200μL，与上述500μL PBS缓冲液混合。在室温下温育30分钟并轻轻摇动后，通过以10,000rpm离心25分钟除去游离的StrandA，并用pH 7.0的10mM PBS冲洗3次，再悬浮在500μL PBS缓冲液中即得。由于FAM的荧光猝灭作用Aptamer C链上的碳量子点不再显示蓝绿色荧光。
实施例2
一种沙门氏菌的检测和消杀方法，步骤如下：
将实施例1核酸适配体-量子点生物传感器分散在磷酸盐缓冲盐水中，制备成浓度为0.32μM的溶液，将上述分散液添加到食品表面用于检测其中的沙门氏菌，沙门氏菌会因高亲和力作用与Aptamer C链特异性结合，A链就会被菌竞争下来，Aptamer C链上的碳量子点会因失去A链上荧光素FAM的猝灭作用而显示出蓝绿色荧光。通过检测碳量子点的蓝绿色荧光用于测定沙门氏菌的含量；
在显示荧光的区域，利用1.5W/cm2 808nm近红外光进行持续照射；杀菌完成后将食品置于磁场中或者在磁铁的作用下分离出Fe3O4纳米粒子，并将食品表面修饰有FAM的Strand A用水洗掉即可。
灵敏性和特异性
特异性
分别使用沙门氏菌、大肠杆菌、副溶血性弧菌以及空白对照组测试本发明生物传感器的检测特异性：将104CFU/100g的沙门氏菌、大肠杆菌、副溶血性弧菌以及无菌的悬浮液分别以实施例2的方法进行检测，得到的实验结果吸光度如图3所示。
由图3可知，采用本发明的生物传感器检测3种菌结果显示，沙门氏菌悬浮液较空白对照组具有强烈的吸光信号，而大肠杆菌、副溶血性弧菌与空白对照组吸光度相当。由此可见，本发明的生物传感器特异性好。
灵敏性
将状态为薄切片的肉制品先进行紫外杀菌，然后使其在无菌状态下在表面接种混匀的沙门氏菌菌悬液，使肉制品表面具有不同沙门氏菌浓度如10，102，103，104CFU/100g，采用实施例2的检测方法，用于评估测定本发明实施例1生物传感器的灵敏度。由图4和图5可知，随着沙门氏菌浓度的增加，用荧光检测器检测的量子点蓝色荧光强度逐渐上升。当细菌浓度范围为10至104CFU/100g时，本发明生物传感器的检测灵敏度为10CFU/100g，标准曲线为y＝0.182x+0.08，相关系数为0.9874，由此可见，本发明的生物传感器具有良好的线性关系和较高的灵敏度。
杀菌效果
采用实施例2的检测方法检测肉制品的沙门氏菌，如图6所示，第一个近红外杀菌前的柱形图结果所示可显示较强的荧光。
若用未沉积PDA的Fe3O4纳米粒子检测沙门氏菌并在显示荧光区域进行近红外杀菌，杀菌完成后将肉制品置于磁场中或者在磁铁的作用下分离出连接有Aptamer C(修饰着量子点)的Fe3O4纳米粒子，并将食品表面修饰有FAM的Strand A用水洗掉，检测杀菌效果，如图6第二个柱形图所示可发现荧光强度在一定程度上变弱。
若用沉积PDA的Fe3O4纳米粒子检测沙门氏菌并在显示荧光区域进行近红外杀菌，杀菌完成后将肉制品置于磁场中或者在磁铁的作用下分离出连接有Aptamer C(修饰着量子点)的Fe3O4纳米粒子，并将食品表面修饰有FAM的Strand A用水洗掉，检测杀菌效果，如图6第三个柱形图所示近红外杀菌后的柱形图所示显示微弱荧光。
表明本发明具有良好的杀菌效果，且PDA引起的光热效应能够增强杀菌效果。
以上所述，仅是本发明的较佳实施例而已，并非是对本发明作其它形式的限制，任何熟悉本专业的技术人员可能利用上述揭示的技术内容加以变更或改型为等同变化的等效实施例。本领域的技术人员可以将其应用于其他致病菌及毒素的检测和杀灭。但是凡是未脱离本发明技术方案内容，依据本发明的技术实质对以上实施例所作的任何简单修改、等同变化与改型，仍属于本发明技术方案的保护范围。
序列表
<110> 青岛农业大学
<120> 用于沙门氏菌检测和消杀的核酸适配体-量子点生物传感器、其制备方法及应用
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 28
<212> DNA
<213> 人工序列(salmonella)
<400> 1
agcagcacag gcgatccaag cttcttca 28
<210> 2
<211> 18
<212> DNA
<213> 人工序列(salmonella)
<400> 2
cgctaggttc gaagaagt 18
Claims (6)
1.用于沙门氏菌检测和消杀的核酸适配体-量子点生物传感器，其特征在于，在沉积有PDA涂层的Fe3O4纳米粒子表面连接经量子点标记的沙门氏菌核酸适配体Aptamer C；并将修饰有荧光素的互补链Strand A与上述Aptamer C配对；
所述沙门氏菌核酸适配体Aptamer C的核苷酸序列如SEQ ID NO.1所示；
所述互补链Strand A的核苷酸序列如SEQ ID NO.2所示；
所述荧光素为FAM或者FITC；
所述的量子点为碳量子点、石墨烯量子点、核壳类和复合材料的无毒荧光量子点中的任意一种。
2.权利要求1所述用于沙门氏菌检测和消杀的核酸适配体-量子点生物传感器的制备方法，其特征在于，步骤如下：
(1)将经量子点标记的Aptamer C链活化并与沉积有PDA涂层的Fe3O4纳米粒子连接：取160μL 2μM的量子点标记的Aptamer C链与30μL 10mg/L的NHS和30μL 10mg/L的EDC混合进行活化；将上述活化的量子点标记的Aptamer C链与沉积有PDA涂层的Fe3O4纳米粒子以体积比为3∶1的比例混合；在室温下温育30分钟并轻轻摇动后，以10,000rpm离心25分钟除去游离的Aptamer C链，并用pH 7.0的10mM PBS冲洗3次，将获得的结合物重新分散在500μL PBS缓冲液中；
(2)将修饰有荧光素的互补链Strand A通过碱基互补配对作用借助氢键与步骤(1)获得的结合物的Aptamer C链进行结合：将200μL 2μM的Strand A加入步骤(1)结合物分散的500μL PBS缓冲液中混合；在室温下温育30分钟并轻轻摇动后，以10,000rpm离心25分钟除去游离的Strand A，并用pH 7.0的10mM PBS冲洗3次，即得。
3.权利要求2所述方法制备的核酸适配体-量子点生物传感器在食品中沙门氏菌检测和消杀中的应用。
4.根据权利要求3所述的应用，其特征在于，所述食品为固相食品。
5.使用权利要求1所述核酸适配体-量子点生物传感器检测和消杀食品中沙门氏菌的方法，其特征在于，步骤如下：
将权利要求1所述核酸适配体-量子点生物传感器分散在磷酸盐缓冲盐水中，制备成浓度为0.32μM的溶液，将上述分散液添加到食品表面用于检测其中的沙门氏菌，沙门氏菌因高亲和力作用与Aptamer C链特异性结合，Strand A被菌竞争下来，Aptamer C链上的量子点会因失去A链上荧光素的猝灭作用而显示出荧光，通过检测量子点的荧光用于测定沙门氏菌的含量；
在量子点显示的荧光区域，利用1.5W/cm2808nm近红外光进行持续照射；杀菌完成后将食品置于磁场中或者在磁铁的作用下分离出Fe3O4纳米粒子，并将食品表面修饰有荧光素的Strand A用水洗掉即可。
6.根据权利要求4所述的应用，其特征在于，所述固相食品为肉制品、果蔬制品中的一种。
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