技术领域technical field
本发明属于检测试剂领域，尤其涉及一种含有双酚A免疫探针的双酚A免疫试纸条及其应用。The invention belongs to the field of detection reagents, in particular to a bisphenol A immune test strip containing a bisphenol A immune probe and an application thereof.
背景技术Background technique
双酚A，也称BPA，在工业上双酚A被用来合成聚碳酸酯(PC)和环氧树脂等材料。60年代以来就被用于制造塑料(奶)瓶、幼儿用的吸口杯、食品和饮料(奶粉)罐内侧涂层。BPA无处不在，从矿泉水瓶、医疗器械到食品包装的内里，都有它的身影。每年，全世界生产2700万吨含有BPA的塑料。但是，BPA也能导致内分泌失调，威胁着胎儿和儿童的健康。癌症和新陈代谢紊乱导致的肥胖也被认为与此有关。欧盟认为含双酚A奶瓶会诱发性早熟，从2011年3月2日起，禁止生产含化学物质双酚A(BPA)的婴儿奶瓶。因此，对于双酚A的检测显得尤为重要。Bisphenol A, also known as BPA, is used industrially to synthesize materials such as polycarbonate (PC) and epoxy resin. Since the 1960s, it has been used to make plastic (milk) bottles, suction cups for children, and coating the inside of food and beverage (milk powder) cans. BPA is everywhere, from mineral water bottles, medical equipment to the inside of food packaging. Every year, 27 million tons of plastic containing BPA are produced worldwide. However, BPA can also lead to endocrine disorders, threatening the health of fetuses and children. Obesity from cancer and metabolic disorders is also thought to be involved. The European Union believes that feeding bottles containing bisphenol A can induce precocious puberty, and since March 2, 2011, the production of baby feeding bottles containing the chemical substance bisphenol A (BPA) has been banned. Therefore, the detection of bisphenol A is particularly important.
现有技术中，对于双酚A的检测方法，经常会受到结构类似的物质的干扰，如：2～氨基苯酚、苯酚、间甲酚等，主要存在着检测特异性差的技术缺陷，无法实现双酚A的准确检测。In the prior art, the detection method of bisphenol A is often interfered by substances with similar structures, such as: 2-aminophenol, phenol, m-cresol, etc., mainly has the technical defect of poor detection specificity, and cannot realize bisphenol A. Accurate detection of phenol A.
因此，研发出一种含有双酚A免疫探针的双酚A免疫试纸条及其应用，用于解决现有技术中，双酚A的检测方法主要存在着检测特异性差、无法实现双酚A的准确检测的技术缺陷，成为了本领域技术人员亟待解决的问题。Therefore, a bisphenol A immune test strip containing a bisphenol A immune probe and its application have been developed to solve the problems of poor detection specificity and inability to achieve bisphenol A detection methods in the prior art. The technical defect of accurate detection of A has become a problem to be solved urgently by those skilled in the art.
发明内容Contents of the invention
有鉴于此，本发明提供了一种含有双酚A免疫探针的双酚A免疫试纸条及其应用，用于解决现有技术中，双酚A的检测方法主要存在着检测特异性差、无法实现双酚A的准确检测的技术缺陷。In view of this, the present invention provides a bisphenol A immunological test strip containing a bisphenol A immune probe and its application, which is used to solve the problems of poor detection specificity, The technical defect that the accurate detection of bisphenol A cannot be realized.
本发明提供了一种双酚A免疫探针的制备方法，所述制备方法为：The present invention provides a kind of preparation method of bisphenol A immune probe, described preparation method is:
步骤一、氯金酸水溶液加热后加入柠檬酸钠水溶液，保温搅拌后，继续搅拌冷却至室温，得第一产物；Step 1. Add sodium citrate aqueous solution after heating the chloroauric acid aqueous solution, after insulated and stirred, continue to stir and cool to room temperature to obtain the first product;
步骤二、所述第一产物调pH至8～9，与对巯基苯甲酸水溶液混合后，震荡摇匀，得第二产物；Step 2. Adjust the pH of the first product to 8-9, mix it with an aqueous solution of p-mercaptobenzoic acid, shake it well, and obtain the second product;
步骤三、所述第二产物与双酚A单克隆抗体溶液混合后，第一次反应，得中间产物；所述中间产物与BSA/PBS溶液混合，第二次反应，离心后去除上清液，滴加去离子水分散，得产品。Step 3, after the second product is mixed with the bisphenol A monoclonal antibody solution, the first reaction is performed to obtain an intermediate product; the intermediate product is mixed with BSA/PBS solution, the second reaction is performed, and the supernatant is removed after centrifugation , dropwise added deionized water to disperse to obtain the product.
优选地，步骤一所述加热的温度为110～120℃，步骤一所述保温搅拌的时间为15min。Preferably, the heating temperature in step 1 is 110-120° C., and the time for heat preservation and stirring in step 1 is 15 minutes.
优选地，所述氯金酸水溶液的浓度为1.0×10-4g/mL，所述柠檬酸钠水溶液的浓度为1.0×10-2g/mL，所述氯金酸水溶液与所述柠檬酸钠水溶液的体积比为100：(0.3～2)。Preferably, the concentration of the aqueous chloroauric acid solution is 1.0×10-4g/mL, the concentration of the aqueous sodium citrate solution is 1.0×10-2 g/mL, the aqueous solution of chloroauric acid and the sodium citrate The volume ratio of the aqueous solution is 100: (0.3-2).
优选地，步骤二中所述第一产物与所述对巯基苯甲酸水溶液的体积比为500：(0.2～1)，所述震荡摇匀的时间为1h。Preferably, the volume ratio of the first product to the aqueous solution of p-mercaptobenzoic acid in step 2 is 500: (0.2-1), and the shaking time is 1 h.
优选地，所述双酚A单克隆抗体的浓度为4～40μg/mL，步骤三中所述第二产物与所述双酚A单克隆抗体溶液的体积比为500：(1～5)，所述第一次反应的时间为1h。Preferably, the concentration of the bisphenol A monoclonal antibody is 4-40 μg/mL, and the volume ratio of the second product to the bisphenol A monoclonal antibody solution in step 3 is 500:(1-5), The time of the first reaction is 1h.
优选地，所述BSA/PBS溶液的浓度为1～3％，所述中间体与所述BSA/PBS溶液的体积比为100：(2～5)。Preferably, the concentration of the BSA/PBS solution is 1-3%, and the volume ratio of the intermediate to the BSA/PBS solution is 100:(2-5).
优选地，所述第一次反应的时间为0.5～1.5h，所述第二次反应的时间为0.8～1.2h。Preferably, the time for the first reaction is 0.5-1.5 h, and the time for the second reaction is 0.8-1.2 h.
优选地，所述离心的转速为7000～12000r/min，所述离心的时间为30min。Preferably, the rotational speed of the centrifugation is 7000-12000 r/min, and the centrifugation time is 30 minutes.
本发明还提供了一种含有以上任意一项所述的制备方法得到的双酚A免疫探针的双酚A免疫试纸条。The present invention also provides a bisphenol A immune test strip containing the bisphenol A immune probe obtained by any one of the above preparation methods.
本发明还提供了以上任意一项所述的制备方法得到的双酚A免疫探针或上述双酚A免疫试纸条在双酚A检测中的应用。The present invention also provides the application of the bisphenol A immune probe obtained by any one of the above preparation methods or the above bisphenol A immune test strip in the detection of bisphenol A.
综上所述，本发明提供了一种双酚A免疫探针的制备方法，本发明还提供了一种含有上述制备方法得到的双酚A免疫探针的双酚A免疫试纸条，本发明还提供了一种上述双酚A免疫探针或双酚A免疫试纸条在双酚A检测中的应用。根据实验测定可得，通过本发明提供的技术方案，对2-氨基苯酚、苯酚、间甲酚、邻甲酚、苯、对苯二酚、双酚A分别进行免疫层析，通过拉曼光谱检测，除双酚A外，均不与双酚A抗体竞争结合，不存在交叉反应，表明本发明提供的技术方案具有高度特异性。解决了现有技术中，双酚A的检测方法主要存在着检测特异性差、无法实现双酚A的准确检测的技术缺陷。同时，本发明提供的技术方案，还具有检测简单、灵敏度高的优点，不仅可以实现双酚A的定性检测，还可以实现双酚A的定量检测。In summary, the present invention provides a method for preparing a bisphenol A immune probe, and the present invention also provides a bisphenol A immune test strip containing the bisphenol A immune probe obtained by the above preparation method. The invention also provides an application of the bisphenol A immune probe or the bisphenol A immune test strip in the detection of bisphenol A. According to the experimental determination, by the technical scheme provided by the present invention, 2-aminophenol, phenol, m-cresol, o-cresol, benzene, hydroquinone, bisphenol A are subjected to immunochromatography respectively, and Raman spectrum In the detection, except for bisphenol A, none compete with the bisphenol A antibody for binding, and there is no cross reaction, indicating that the technical solution provided by the present invention has high specificity. The method solves the technical defects of poor detection specificity and inability to realize accurate detection of bisphenol A mainly in the detection method of bisphenol A in the prior art. At the same time, the technical solution provided by the invention also has the advantages of simple detection and high sensitivity, and can not only realize the qualitative detection of bisphenol A, but also realize the quantitative detection of bisphenol A.
附图说明Description of drawings
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据提供的附图获得其他的附图。In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only It is an embodiment of the present invention, and those skilled in the art can also obtain other drawings according to the provided drawings on the premise of not paying creative efforts.
图1为本发明提供的一种双酚A免疫试纸条的结构示意图；Fig. 1 is the structural representation of a kind of bisphenol A immune test strip provided by the present invention;
图2为本发明提供的一种双酚A免疫探针的紫外-可见吸收光谱图；Fig. 2 is the ultraviolet-visible absorption spectrogram of a kind of bisphenol A immunoprobe provided by the present invention;
图3为本发明提供的一种双酚A检测方法的双酚A标准曲线图；Fig. 3 is the bisphenol A standard curve figure of a kind of bisphenol A detection method provided by the present invention;
图4为本发明提供的一种双酚A检测方法的重复性考察结果图。Fig. 4 is a diagram of the repeatability investigation results of a bisphenol A detection method provided by the present invention.
具体实施方式detailed description
本发明提供了一种含有双酚A免疫探针的双酚A免疫试纸条及其应用，用于解决现有技术中，双酚A的检测方法主要存在着检测特异性差、无法实现双酚A的准确检测的技术缺陷。The invention provides a bisphenol A immune test strip containing a bisphenol A immune probe and its application, which is used to solve the problems of poor detection specificity and inability to realize bisphenol A detection methods in the prior art. A's accurate detection of technical flaws.
下面将对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some of the embodiments of the present invention, but not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.
为了更详细说明本发明，下面结合实施例对本发明提供的一种含有双酚A免疫探针的双酚A免疫试纸条及其应用，进行具体地描述。In order to describe the present invention in detail, a bisphenol A immunological test strip containing a bisphenol A immunological probe provided by the present invention and its application will be specifically described below in conjunction with the examples.
实施例1Example 1
本实施例为根据本发明提供的双酚A免疫探针的制备方法，结合本领域技术人员都熟知的试纸条制备方法，制备双酚A免疫试纸条的具体实施例。This example is a specific example of preparing a bisphenol A immune test strip according to the preparation method of the bisphenol A immune probe provided by the present invention, combined with the test strip preparation method well known to those skilled in the art.
1、制备胶体金1. Preparation of colloidal gold
在剧烈搅拌下，将200mL 1.0×10-4g·mL-1的氯金酸水溶液110℃～120℃油浴加热，迅速加入1.8mL 1.0×10-2g·mL-1的柠檬酸钠水溶液，保持沸腾15分钟后自然冷却搅拌至室温，得到胶体金溶液。Under vigorous stirring, heat 200mL of 1.0×10-4 g·mL-1 chloroauric acid aqueous solution in an oil bath at 110℃～120℃, and quickly add 1.8mL of 1.0×10-2 g·mL-1 sodium citrate aqueous solution , kept boiling for 15 minutes and then cooled and stirred to room temperature naturally to obtain a colloidal gold solution.
2、4MBA标记胶体金2. 4MBA labeled colloidal gold
吸取上述制得的胶体金溶液10mL置于离心管中，用碳酸钾溶液将其pH值调为8～9，加入10μL 1mM的对巯基苯甲酸(4MBA)，振荡摇匀，常温下反应1小时。Take 10 mL of the colloidal gold solution prepared above and place it in a centrifuge tube, adjust its pH value to 8-9 with potassium carbonate solution, add 10 μL of 1 mM p-mercaptobenzoic acid (4MBA), shake well, and react at room temperature for 1 hour .
3、制备4MBA标记免疫探针3. Preparation of 4MBA-labeled immunoprobe
向上述制得的4MBA标记胶体金溶液中加入50μL滴度为1:1000的双酚A单克隆抗体，反应1小时后，加入300μL 3％BSA/PBS溶液。反应1小时后，7000r/min高速离心30分钟。去除上层清液，滴加适量去离子水将胶体金分散均匀至1mL，得到4MBA标记胶体金免疫探针。50 μL of bisphenol A monoclonal antibody with a titer of 1:1000 was added to the 4MBA-labeled colloidal gold solution prepared above, and after reacting for 1 hour, 300 μL of 3% BSA/PBS solution was added. After reacting for 1 hour, centrifuge at a high speed of 7000 r/min for 30 minutes. The supernatant was removed, and an appropriate amount of deionized water was added dropwise to disperse the colloidal gold evenly to 1 mL to obtain a 4MBA-labeled colloidal gold immunoprobe.
4、包被NC膜4. Coated with NC film
配置BVA-PLL浓度为1mg/mL，羊抗鼠IgG 0.1mg/mL。通过滑膜喷金仪在NC膜的T线、C线分别取10μL划线，在室温下包被20min，37℃烘箱烘干1h。Configure BVA-PLL concentration as 1mg/mL and goat anti-mouse IgG as 0.1mg/mL. 10 μL was drawn on the T line and C line of the NC film by a synovial film gold sprayer, coated at room temperature for 20 min, and oven-dried at 37 °C for 1 h.
5、组装试纸条5. Assemble the test strips
按照图1的结构，将NC膜贴到PVC板的中间位置，吸水纸和样品垫贴于NC膜的两端，与NC膜重叠大概1.5mm。最后，将组装好的试纸条剪成6mm宽，放置于洁净的塑料盒中4℃贮存备用。According to the structure in Figure 1, stick the NC membrane to the middle of the PVC board, and stick the absorbent paper and the sample pad to the two ends of the NC membrane, overlapping with the NC membrane by about 1.5mm. Finally, the assembled test strips were cut into 6 mm wide, and stored in a clean plastic box at 4°C for later use.
实施例2Example 2
本实施例为标准曲线的制作及检测线试验的具体实施例。This embodiment is the specific embodiment of the making of standard curve and detection line test.
准确称取0.1g双酚A固体溶于1mL乙醇，用去离子水稀释至不同浓度(100、50、30、10、5、0ng/mL)。在组装好的试纸条上，将不同浓度的双酚A溶液滴加在样品垫上，层析过程在15min内完成，T线和C线会出现相应的显色。当待测样品中BPA的浓度大于30ng/mL，则胶体金拉曼标记免疫探针的抗体表位全部被待测样品中BPA结合，故而无法与T线上的固定抗原发生特异性反应，T线不显色，为阴性；当待测样品中BPA的浓度小于30ng/mL，则胶体金拉曼标记免疫探针的抗体表位有剩余，会与T线上的固定抗原发生特异性反应，T线呈紫色，为阳性。Accurately weigh 0.1 g of bisphenol A solid, dissolve it in 1 mL of ethanol, and dilute to different concentrations (100, 50, 30, 10, 5, 0 ng/mL) with deionized water. On the assembled test strip, drop bisphenol A solutions of different concentrations on the sample pad, the chromatography process is completed within 15 minutes, and corresponding color development will appear on the T line and C line. When the concentration of BPA in the sample to be tested is greater than 30ng/mL, the antibody epitopes of the colloidal gold Raman-labeled immunoprobe are all bound by BPA in the sample to be tested, so they cannot specifically react with the fixed antigen on the T line, and the T The line does not develop color and is negative; when the concentration of BPA in the sample to be tested is less than 30ng/mL, the antibody epitope of the colloidal gold Raman-labeled immunoprobe remains, which will specifically react with the fixed antigen on the T line. The T line is purple and positive.
用拉曼光谱仪检测T线区的4MBA的拉曼信号，在最优条件下，以1077cm-1与844cm-1处峰位的峰面积的比值对应双酚A浓度制作标准曲线。得到该方法检测双酚A的线性范围为5～50ng/mL，标准曲线方程为y＝-0.0472x+2.8833，R2＝0.992，最低检测线为0.5ng/mL。Use a Raman spectrometer to detect the Raman signal of 4MBA in the T line region. Under optimal conditions, use the ratio of the peak area at 1077cm-1 to the peak at 844cm-1 to make a standard curve corresponding to the concentration of bisphenol A. The linear range of detecting bisphenol A by this method is 5-50 ng/mL, the equation of the standard curve is y=-0.0472x+2.8833, R2 =0.992, and the lowest detection line is 0.5 ng/mL.
实施例3Example 3
本实施例为特异性、信号均一性、重复性和添加回收实验的具体实施例。This example is a specific example of specificity, signal uniformity, repeatability and addition recovery experiments.
3.1特异性考察3.1 Specificity investigation
取100μL的50ng/mL的2-氨基苯酚、苯酚、间甲酚、邻甲酚、苯、对苯二酚、双酚A和0ng/mL双酚A溶液分别滴加在试纸条上，进行免疫层析。通过拉曼光谱检测，所有药物不与双酚A抗体竞争结合，不存在交叉反应，表明该基于表面增强拉曼光谱结合胶体金免疫层析检测双酚A的方法具有高度特异性。Take 100 μL of 50ng/mL 2-aminophenol, phenol, m-cresol, o-cresol, benzene, hydroquinone, bisphenol A and 0ng/mL bisphenol A solution and add them dropwise on the test strip, and carry out Immunochromatography. Through Raman spectroscopy detection, all drugs did not compete with the bisphenol A antibody for binding, and there was no cross-reaction, indicating that the method based on surface-enhanced Raman spectroscopy combined with colloidal gold immunochromatography for the detection of bisphenol A was highly specific.
3.2信号均一性考察3.2 Inspection of Signal Uniformity
取100μL的100ng/mL的双酚A溶液滴加在试纸条上，进行免疫层析。任意选取10个点进行检测，以1077cm-1与844cm-1处峰位的峰面积的比值作为判断标准，结果如表1所示。Take 100 μL of 100 ng/mL bisphenol A solution and drop it on the test strip for immunochromatography. Ten points were arbitrarily selected for detection, and the ratio of the peak areas at 1077 cm-1 to 844 cm-1 was used as the judgment standard. The results are shown in Table 1.
表1Table 1
表1结果表明，该免疫层析试纸条具有很高的信号均一性，检测精密度较高。The results in Table 1 show that the immunochromatographic test strip has high signal uniformity and high detection precision.
3.3重复性考察3.3 Repeatability inspection
分别取100μL0和50ng/mL的双酚A溶液滴加在试纸条上，进行免疫层析。通过拉曼光谱仪检测信号，间隔两周后重复检测。结果表明，两次检测数据基本相同，即该基于表面增强拉曼光谱结合胶体金免疫层析检测双酚A的方法重复性良好。具体实验结果可参阅附图4。附图4中，由上至下四条曲线，分别为a、b、c、d，其中，a和b分别为0ng/mLBPA间隔两周的拉曼图谱，b和d分别为50ng/mLBPA间隔两周的拉曼图谱。Take 100 μL of 0 and 50 ng/mL bisphenol A solutions and drop them on the test strips for immunochromatography. Signals were detected by Raman spectroscopy and repeated at two week intervals. The results showed that the two detection data were basically the same, that is, the method based on surface-enhanced Raman spectroscopy combined with colloidal gold immunochromatography for the detection of bisphenol A had good repeatability. Please refer to accompanying drawing 4 for specific experimental results. In accompanying drawing 4, four curves from top to bottom are respectively a, b, c, and d, wherein, a and b are the Raman spectra of 0 ng/mLBPA at intervals of two weeks respectively, and b and d are the Raman spectra of 50 ng/mLBPA at intervals of two weeks, respectively. Zhou's Raman spectrum.
3.4样品添加回收实验3.4 Sample addition recovery experiment
为了测定该方法在实际检测中的可靠性和准确性，将不同浓度的双酚A溶液加标到四种环境水样中。将珠江水、桶装水、怡宝水和自来水先用0.45μm的微孔滤膜抽滤后，对每个样品添加双酚A标准溶液至浓度为5、10、50ng/mL。分别滴在试纸条上，进行免疫层析。采集拉曼信号，每个样品重复测量3次，取平均值。所得实验结果可参阅表2。In order to determine the reliability and accuracy of the method in actual detection, different concentrations of bisphenol A solutions were spiked into four environmental water samples. After the Pearl River water, bottled water, C’estbon water and tap water were filtered through a 0.45 μm microporous membrane, each sample was added with bisphenol A standard solution to a concentration of 5, 10 and 50 ng/mL. They were dropped on test strips for immunochromatography. The Raman signal was collected, and the measurement was repeated three times for each sample, and the average value was taken. The experimental results obtained can be referred to Table 2.
表2Table 2
表2提供的结果表明，建立的基于表面增强拉曼光谱结合胶体金免疫层析检测双酚A的方法能够满足检测要求，可用于实际水样中双酚A残留的分析检测。The results provided in Table 2 show that the established method based on surface-enhanced Raman spectroscopy combined with colloidal gold immunochromatography for the detection of bisphenol A can meet the detection requirements and can be used for the analysis and detection of bisphenol A residues in actual water samples.
综上所述，本发明提供了一种双酚A免疫探针的制备方法，本发明还提供了一种含有上述制备方法得到的双酚A免疫探针的双酚A免疫试纸条，本发明还提供了一种上述双酚A免疫探针或双酚A免疫试纸条在双酚A检测中的应用。根据实验测定可得，通过本发明提供的技术方案，对2-氨基苯酚、苯酚、间甲酚、邻甲酚、苯、对苯二酚、双酚A分别进行免疫层析，通过拉曼光谱检测，除双酚A外，均不与双酚A抗体竞争结合，不存在交叉反应，表明本发明提供的技术方案具有高度特异性。解决了现有技术中，双酚A的检测方法主要存在着检测特异性差、无法实现双酚A的准确检测的技术缺陷。同时，本发明提供的技术方案，还具有检测简单、灵敏度高的优点，不仅可以实现双酚A的定性检测，还可以实现双酚A的定量检测。In summary, the present invention provides a method for preparing a bisphenol A immune probe, and the present invention also provides a bisphenol A immune test strip containing the bisphenol A immune probe obtained by the above preparation method. The invention also provides an application of the bisphenol A immune probe or the bisphenol A immune test strip in the detection of bisphenol A. According to the experimental determination, by the technical scheme provided by the present invention, 2-aminophenol, phenol, m-cresol, o-cresol, benzene, hydroquinone, bisphenol A are subjected to immunochromatography respectively, and Raman spectrum In the detection, except for bisphenol A, none compete with the bisphenol A antibody for binding, and there is no cross reaction, indicating that the technical solution provided by the present invention has high specificity. The method solves the technical defects of poor detection specificity and inability to realize accurate detection of bisphenol A mainly in the detection method of bisphenol A in the prior art. At the same time, the technical solution provided by the invention also has the advantages of simple detection and high sensitivity, and can not only realize the qualitative detection of bisphenol A, but also realize the quantitative detection of bisphenol A.
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。The above is only a preferred embodiment of the present invention, it should be pointed out that, for those of ordinary skill in the art, without departing from the principle of the present invention, some improvements and modifications can also be made, and these improvements and modifications can also be made. It should be regarded as the protection scope of the present invention.
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610531732.2ACN106153928A (en) | 2016-07-06 | 2016-07-06 | A kind of bisphenol-A immunity test strip containing bisphenol-A immunological probe and application thereof |
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610531732.2ACN106153928A (en) | 2016-07-06 | 2016-07-06 | A kind of bisphenol-A immunity test strip containing bisphenol-A immunological probe and application thereof |
| Publication Number | Publication Date |
|---|---|
| CN106153928Atrue CN106153928A (en) | 2016-11-23 |
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610531732.2APendingCN106153928A (en) | 2016-07-06 | 2016-07-06 | A kind of bisphenol-A immunity test strip containing bisphenol-A immunological probe and application thereof |
| Country | Link |
|---|---|
| CN (1) | CN106153928A (en) |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109632764A (en)* | 2019-01-07 | 2019-04-16 | 辽宁石油化工大学 | Preparation method and application of a flexible SERS sensor for detecting bisphenol A |
| CN110133255A (en)* | 2018-02-02 | 2019-08-16 | 中国人民解放军军事科学院军事医学研究院 | A rapid and highly sensitive SERS-immunochromatographic detection method for the detection of Mycoplasma pneumoniae infection |
| CN110133254A (en)* | 2018-02-02 | 2019-08-16 | 中国人民解放军军事科学院军事医学研究院 | A plug-in type Raman chromatography test strip that can realize signal secondary amplification |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102221542A (en)* | 2011-03-24 | 2011-10-19 | 华南师范大学 | Method for detecting Clenbuterol by applying competitive SERS (Surface-Enhanced Raman Scattering) and application thereof |
| CN103048445A (en)* | 2012-12-14 | 2013-04-17 | 广东工业大学 | Preparation method of bisphenol A-coated antigen with polylysine as carrier and application thereof |
| CN105259158A (en)* | 2015-11-13 | 2016-01-20 | 暨南大学 | Surface enhanced Raman scattering immunochromatography test paper strip and preparation method and application |
| CN105352930A (en)* | 2015-12-01 | 2016-02-24 | 深圳市绿诗源生物技术有限公司 | Bisphenol A fluorescence polarization immunity analysis and detection method based on adopting AMF as fluorescein marker |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102221542A (en)* | 2011-03-24 | 2011-10-19 | 华南师范大学 | Method for detecting Clenbuterol by applying competitive SERS (Surface-Enhanced Raman Scattering) and application thereof |
| CN103048445A (en)* | 2012-12-14 | 2013-04-17 | 广东工业大学 | Preparation method of bisphenol A-coated antigen with polylysine as carrier and application thereof |
| CN105259158A (en)* | 2015-11-13 | 2016-01-20 | 暨南大学 | Surface enhanced Raman scattering immunochromatography test paper strip and preparation method and application |
| CN105352930A (en)* | 2015-12-01 | 2016-02-24 | 深圳市绿诗源生物技术有限公司 | Bisphenol A fluorescence polarization immunity analysis and detection method based on adopting AMF as fluorescein marker |
| Title |
|---|
| PINONG HUANG ET AL: "A fluorescence polarization immunoassay method for detection of the bisphenol A residue in environmental water samples based on a monoclonal antibody and 4′-(aminomethyl)fluorescein", 《ANALYTICAL METHODS》* |
| 李明心: "基于贵金属纳米颗粒的表面増强拉曼散射免疫层析分析方法的研究", 《中国优秀硕士论文全文数据库工程科技I辑》* |
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110133255A (en)* | 2018-02-02 | 2019-08-16 | 中国人民解放军军事科学院军事医学研究院 | A rapid and highly sensitive SERS-immunochromatographic detection method for the detection of Mycoplasma pneumoniae infection |
| CN110133254A (en)* | 2018-02-02 | 2019-08-16 | 中国人民解放军军事科学院军事医学研究院 | A plug-in type Raman chromatography test strip that can realize signal secondary amplification |
| CN109632764A (en)* | 2019-01-07 | 2019-04-16 | 辽宁石油化工大学 | Preparation method and application of a flexible SERS sensor for detecting bisphenol A |
| Publication | Publication Date | Title |
|---|---|---|
| Krska et al. | Rapid test strips for analysis of mycotoxins in food and feed | |
| Sun et al. | One-step time-resolved fluorescence microsphere immunochromatographic test strip for quantitative and simultaneous detection of DON and ZEN | |
| CN103018467B (en) | Colloidal-gold detecting-card of bisphenol-A and preparation method thereof | |
| Chon et al. | Simultaneous immunoassay for the detection of two lung cancer markers using functionalized SERS nanoprobes | |
| Li et al. | On-site ultrasensitive detection paper for multiclass chemical contaminants via universal bridge-antibody labeling: Mycotoxin and illegal additives in milk as an example | |
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