Allergen detection sensor, preparation method thereof and rapid detection device

Abstract

The invention provides an allergen detection sensor, a preparation method thereof and a rapid detection device, the sensor comprises a response signal material, the response signal material is a carbon quantum dot material prepared based on plant cocklebur fruit, and the carbon quantum dot material has specific recognition capability on a target allergen; the reference signal material is combined with the carbon quantum dot material to form a proportional fluorescent probe; and the carrier is loaded with a proportional fluorescent probe. The sensor provided by the invention has the characteristics of simple preparation, low cost, rapid reaction and high specificity, and can be used for rapid detection of allergens.

Description

Technical Field

[0001] This application relates to the field of allergen detection methods, and in particular to an allergen detection sensor, its preparation method, and a rapid detection device. Background Technology

[0002] Studies have shown that wormwood pollen is one of the main sources of pollen allergies. In Europe, 10%–14% of hay fever sufferers are allergic to common wormwood pollen. In northern regions, wormwood pollen accounts for over 60% of autumn hay fever sufferers, with an annual incidence rate increasing at 7.2%. Direct medical expenses related to wormwood pollen allergy in China exceed 2 billion yuan annually, and lost work / school time continues to rise. Artv1 is a major allergen in wormwood pollen and a key factor in inducing immune responses in sensitive individuals. Due to its central role in wormwood pollen allergy, Artv1 has become an important research target for the diagnosis and treatment of wormwood pollen allergy.

[0003] There are two common methods for detecting pollen allergens in clinical practice: one is antibody-based methods, including enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). These two molecular diagnostic techniques are commonly used in immunology, determining the allergen based on the concentration of the corresponding antibody (sIgE). However, these methods are relatively cumbersome and require the use of antibodies twice, leading to high testing costs. Furthermore, these methods rely on large-scale instruments, hindering their widespread adoption. The other method is antigen-based skin prick testing (SPT). While SPT is fast and convenient, it relies on the operator's judgment of the spot size, and the accuracy of the results is particularly affected by the actual allergen content in the diagnostic reagent and cross-allergy, resulting in a false positive rate of over 20%. In addition, SPT can cause allergic reactions in some patients. Summary of the Invention

[0004] To address the aforementioned problems, this invention provides an allergen detection sensor, its preparation method, and a rapid detection device, thereby solving the issues of existing wormwood pollen allergy detection technologies being time-consuming, reliant on large equipment, expensive, and inaccurate.

[0005] In a first aspect, the present invention provides an allergen detection sensor, the technical solution of which is: An allergen detection sensor, the sensor comprising: The response signal material is a carbon quantum dot material prepared based on the plant Xanthium sibiricum, and the carbon quantum dot material has the ability to specifically recognize the target allergen. A reference signal material, wherein the reference signal material is combined with the carbon quantum dot material to form a proportional fluorescent probe; The carrier on which the proportional fluorescent probe is loaded.

[0006] As one preferred embodiment, the reference signal material includes one or more of europium trichloride hexahydrate, rhodamine B, platinum porphyrin, tridichloro-tris(2,2'-bipyridine)ruthenium(II) hexahydrate, dichloro-tris(1,10-phenanthroline)ruthenium(II) tetrahydrate, dichloro-bis(2,2'-bipyridine)(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dihydrate, and isothiocyanate (FITC).

[0007] As one preferred embodiment, the carrier includes one or more of nonwoven fabric, spunbond fabric, filter paper, nitrocellulose membrane, and polyvinylidene fluoride membrane.

[0008] Secondly, the present invention provides a method for preparing an allergen detection sensor, the technical solution of which is: A method for preparing an allergen detection sensor, the method comprising: The response signal material is prepared into a first solution of a certain concentration by adding ultrapure water; wherein, the response signal material is a carbon quantum dot material prepared based on the plant Xanthium sibiricum, and the carbon quantum dot material has the ability to specifically recognize the target allergen; The reference signal material is dissolved in a solvent to prepare a second solution of a certain concentration; Under a nitrogen atmosphere, the first solution and the second solution are mixed in a certain proportion and stirred at room temperature for a certain time to combine the carbon quantum dot material and the response signal material. After the reaction was completed, the reaction system was transferred to a dialysis bag and dialyzed with excess ultrapure water for a certain period of time to obtain a proportional fluorescent probe. The proportional fluorescent probe is loaded onto a carrier to prepare an allergen detection sensor.

[0009] As one of the preferred methods, the loading method for loading the proportional fluorescent probe onto the carrier includes one or more of the following: impregnation, electrostatic binding, and adsorption.

[0010] As one of the preferred options, the solvent includes any one of methanol, acetonitrile, ultrapure water, and ethanol.

[0011] As one of the preferred embodiments, the concentration of the first solution is 0.001 mg / mL to 3 mg / mL; the ratio of the first solution to the second solution is 10:1 to 1:10; the stirring time is 1 h to 5 h; and the stirring speed is 100 rpm to 500 rpm.

[0012] As one of the preferred solutions, the loading time for the proportional fluorescent probe onto the carrier is 1h to 5h, and the loading temperature is 5℃ to 45℃.

[0013] Thirdly, this invention provides a rapid allergen detection device, the technical solution of which is: A rapid allergen detection device, comprising: Allergen detection sensor, such as the allergen detection sensor provided in the first aspect of the present invention; The dark box has notches in the side walls and a light-transmitting window at the bottom. The support body enters the dark box through the notch and is sealed and snapped onto the notch wall of the dark box; the support body has a slot in which the allergen detection sensor is placed. The light source is located on the top wall of the darkroom; The camera device is located below the bottom wall of the darkroom; The camera lens, the light source, the light-transmitting window, and the allergen detection sensor are positioned relative to each other.

[0014] As one preferred embodiment, the camera device includes a mobile phone holder and a mobile phone, with the mobile phone holder positioned below the dark box and the SIM card attached to the mobile phone holder; The mobile phone has a built-in camera and image processing unit; wherein... The lens is used to capture the fluorescence image generated by the allergen detection sensor when it comes into contact with the target allergen under the excitation of the light source; The image processing unit is used to convert the colors in the fluorescence image into RGB values ​​so as to obtain the concentration of the target allergen by combining it with a standard curve; the standard curve represents the trend of the RGB values ​​of the target allergen at different concentrations.

[0015] Artemisia pollen allergy is a common symptom in northern my country. There are generally three detection methods: 1. Skin prick test. This test is simple, quick, and inexpensive, making it a commonly used clinical screening method. However, antihistamines must be discontinued before testing, and it is not suitable for those with severe skin allergies or skin diseases. Results rely on experience and there is a probability of false positives. 2. Serum-specific IgE test. This test does not require discontinuation of antihistamines and is safer for those with sensitive skin or those unsuitable for skin tests. The results are objective and quantifiable. However, the testing process is more complex, using expensive antibodies and large instruments, making it unsuitable for widespread use. 3. Nasal mucosal provocation test. This test more closely resembles the natural allergic state and has high specificity, but it is relatively complex to perform and may cause patient discomfort, thus its clinical application is less common.

[0016] Compared with the above detection methods, the allergen detection sensor provided by the present invention has the following advantages: In this embodiment of the invention, the carbon quantum dot combined with proportional fluorescent probe technology used in the allergen detection sensor exhibits stable performance and can be stored long-term on a specific carrier. The specific binding groups of carbon quantum dots and the proportional fluorescent probe technology enable rapid response to wormwood pollen allergens, with a short reaction time; detection results can be obtained in just 3 minutes. The allergen detection sensor provided in this embodiment of the invention has a simple preparation process, and the raw material, cocklebur, is cheap and readily available with low storage costs. Therefore, using this sensor for the detection of wormwood pollen allergens can greatly reduce detection costs and shorten the detection time. The allergen detection sensor provided in this invention exhibits excellent specificity when used as a tool for detecting wormwood pollen allergens. The combination of carbon quantum dot preparation and proportional fluorescence probe technology is based on the specific recognition of wormwood pollen allergens. Therefore, during detection, it shows extremely low interference response to other pollens or substances, achieving good detection results for major wormwood pollen allergens with a specificity exceeding 95%.

[0017] The method and apparatus described herein have the same advantages over the prior art as the aforementioned sensors, and will not be repeated here. Attached Figure Description

[0018] To more clearly illustrate the technical solution of this application, the drawings used in the description of this application will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 This is a flowchart illustrating the steps of a method for preparing an allergen detection sensor according to an embodiment of the present invention. Figure 2 The fluorescence spectrum of an allergen detection sensor provided in an embodiment of the present invention is shown. Figure 3 This is a graph showing the detection of Artv1 protein by an allergen detection sensor provided in an embodiment of the present invention at different times; Figure 4 This is a diagram illustrating the detection of different proteins in plasma by an allergen detection sensor provided in an embodiment of the present invention. Figure 5 This is a schematic diagram of the structure of a rapid allergen detection device provided in an embodiment of the present invention; Figure 6 This is a schematic diagram illustrating the method of using the rapid allergen detection device provided in an embodiment of the present invention; Figure 7 This is a linear relationship graph of allergen detection using a rapid allergen detection device according to an embodiment of the present invention.

[0020] Explanation of reference numerals in the attached figures: 1. Light source; 2. Dark box; 3. Support structure; 4. Mobile phone holder; 5. Light-transmitting window. Detailed Implementation

[0021] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit the present invention or its application or use. Based on the embodiments of the present invention, any product that is the same as or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the protection scope of the present invention. Furthermore, all other embodiments obtained by those skilled in the art without inventive effort are within the protection scope of the present invention.

[0022] Specific experimental steps or conditions are not specified in the embodiments; they can be performed according to the conventional experimental steps or conditions described in the prior art. Reagents and other instruments used, unless otherwise specified, are all commercially available conventional reagent products. Furthermore, the accompanying drawings are merely illustrative diagrams of the embodiments of the present invention and are not necessarily drawn to scale. The same reference numerals in the drawings denote the same or similar parts, and therefore, repeated descriptions of them will be omitted. Some block diagrams shown in the drawings are functional entities and do not necessarily correspond to physically or logically independent entities.

[0023] Techniques, methods, and devices known to those skilled in the art may not be discussed in detail, but where appropriate, such techniques, methods, and devices should be considered part of this specification.

[0024] In the description of this invention, it should be understood that the use of terms such as "first" and "second" to define components is merely for the purpose of distinguishing the corresponding components. Unless otherwise stated, the above terms have no special meaning and therefore should not be construed as limiting the scope of protection of this invention.

[0025] Furthermore, the technical features involved in the different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0026] To better understand the technical solution of this application, the functionalized carbon quantum dots, proportional fluorescence probe technology, and rapid detection principle will now be explained: In this invention, functionalized carbon quantum dots prepared from the plant Xanthium sibiricum are cleverly designed to accurately identify Artemisia pollen allergens, much like a "key" specifically adapted to Artemisia pollen allergens, laying the foundation for subsequent detection processes.

[0027] As we know, carbon quantum dot fluorescent probe technology and ratiometric fluorescent probe technology are both cutting-edge optical sensing detection technologies for allergen detection (such as allergenic proteins or molecules in food, air, and drugs). Carbon quantum dots emit fluorescence at specific wavelengths when excited by light. When carbon quantum dots bind to specific substances (such as allergen proteins, antigens, and metal ions), the fluorescence intensity of the carbon dots changes (quenching or enhancement). A ratiometric fluorescent probe is a probe system that emits fluorescence at two or more wavelengths; it reflects the concentration of the target substance by the ratio of emitted light intensities.

[0028] Although carbon quantum dots are highly sensitive, relying solely on changes in their fluorescence signal intensity to detect allergens is insufficient. Factors such as the concentration of the carbon quantum dot probe, the intensity of the light source, the temperature, pH value, and sample turbidity of the detection environment can all affect the fluorescence intensity, leading to inaccurate measurement results. When detecting allergens using a single fluorescence signal, only an increase or decrease in fluorescence intensity (i.e., brightening or darkening) can usually be observed, which is difficult to judge accurately with the naked eye, let alone achieve intuitive color changes. Observation under fluorescence spectroscopy or excitation light is typically required, making portable or rapid visualization difficult.

[0029] Proportional fluorescence probe technology can eliminate interference from environmental factors, making the results more stable and reliable. However, existing probe systems have drawbacks such as high cost of fluorescent signal materials, slow response, difficulty in doping and poor binding stability, and small wavelength spacing between the two emission peaks, resulting in indistinct color changes. Therefore, they are not suitable for rapid on-site visual detection of wormwood pollen allergies.

[0030] In view of this, it is necessary to develop a novel allergen detection method and device that is rapid, accurate, convenient, and low-cost. According to a first aspect of the present invention, an allergen detection sensor is provided, comprising: a response signal material, wherein the response signal material is a carbon quantum dot material prepared based on the plant Xanthium sibiricum, and the carbon quantum dot material has a specific recognition ability for the target allergen; a reference signal material, wherein the reference signal material is combined with the carbon quantum dot material to form a proportional fluorescent probe; and a carrier on which the proportional fluorescent probe is loaded.

[0031] Specifically, the sensor in this embodiment of the invention is prepared by combining carbon quantum dots with proportional fluorescent probe technology, which have specific recognition capabilities for target allergens, and then loading them onto a specific carrier. The target allergen can be a pollen allergen, specifically an artemisia pollen allergen, and more specifically an Artv1 allergen. The carbon quantum dot material is prepared from the plant Xanthium sibiricum, possessing groups that can specifically bind to artemisia pollen allergens. Combined with fluorescence technology, it offers the feasibility of rapid and convenient detection. Xanthium sibiricum is an annual herb belonging to the Asteraceae family. Its fruit, Xanthium sibiricum seeds, is a traditional Chinese medicine used to treat allergic rhinitis. It contains abundant sesquiterpene lactones, caffeoyl xanthocyclin, xanthine, and xanthanyl lactones, among other anti-allergic active substances. These are synthesized into carbon quantum dots via microwave hydrothermal synthesis, retaining the active substances and thus exhibiting specificity for artemisia pollen allergens. The reference signal material can include fluorescent materials, inorganic luminescent materials, and polymeric fluorescent particles, etc. In this embodiment, a fluorescent material is selected as the reference signal material. In this embodiment, carbon quantum dot material is used as the response signal of the proportional fluorescence probe, and the fluorescent material is used as the reference signal of the proportional fluorescence probe, which are combined to form a proportional fluorescence probe.

[0032] By combining proportional fluorescence probe technology with functionalized carbon quantum dots, the emission peak intensity of the carbon quantum dot material changes when the target allergen is present, while the reference signal material maintains a stable signal. When the carbon quantum dots specifically bind to wormwood pollen allergens, the change in the ratio of the two fluorescence emission peak intensities is visually apparent through ultraviolet light irradiation, revealing a color change. This visualized output allows testing personnel to quickly determine the presence of wormwood pollen allergens without complex instrument analysis, significantly improving detection efficiency and convenience. Finally, the proportional fluorescence probe is loaded onto a carrier for long-term storage.

[0033] In this embodiment of the invention, the proportional fluorescent probe using carbon quantum dot materials as the response signal material differs from existing proportional fluorescent probe technologies and has the following advantages: Carbon quantum dot materials exhibit high sensitivity, and the carbon quantum dot-based proportional fluorescent probe technology used in the provided allergen detection sensor demonstrates stable performance and can be stored long-term on specific carriers. The specific binding groups of carbon quantum dots and the proportional fluorescent probe technology enable rapid response to wormwood pollen allergens, with a short reaction time; detection results can be obtained in as little as 3 minutes.

[0034] The preparation steps of the allergen detection sensor are simple, and the raw material, cocklebur, is cheap and readily available with low storage costs. Therefore, using this sensor for the detection of wormwood pollen allergens can greatly reduce detection costs and shorten the detection time, which is conducive to large-scale production and practical application.

[0035] The sufficiently large difference in wavelength between the emission peaks of carbon quantum dot materials and fluorescent materials leads to a more pronounced overall color change upon contact with allergens. This visible color change in the sensor makes result interpretation more intuitive, enabling qualitative screening without instrument assistance. Suitable for rapid on-site detection, it can further be applied to portable sensing applications in the field of biological detection. For example, by lowering the detection threshold, it can serve as a foundation for developing low-cost, portable devices for rapid, home-use allergen detection.

[0036] Carbon quantum dot materials and fluorescent materials are easy to synthesize and have stable and reliable systems, which greatly improves the anti-interference ability and detection accuracy of probes, providing a reliable technical path for developing high-performance allergen detection methods.

[0037] Thus, this embodiment of the invention retains the excellent response characteristics of carbon quantum dots, combined with the optical stability and color controllability of fluorescent materials. The resulting allergen detection sensor exhibits excellent specificity when used as a tool for detecting wormwood pollen allergens. The combination of carbon quantum dot preparation and proportional fluorescence probe technology is based on the specific recognition of wormwood pollen allergens. Therefore, during detection, it exhibits extremely low interference response to other pollens or substances, demonstrating excellent detection performance for major wormwood pollen allergens, with a specificity exceeding 95%. This sensor combines detection sensitivity and specificity with intuitive visual judgment, making it a promising solution for rapid, visualized allergen detection.

[0038] As a further preferred design of this embodiment, the fluorescent material required as the reference signal is preferably one or more of europium trichloride hexahydrate, rhodamine B, platinum porphyrin, tridichloro-tris(2,2'-bipyridine)ruthenium(II) hexahydrate, dichloro-tris(1,10-phenanthroline)ruthenium(II) tetrahydrate, dichloro-bis(2,2'-bipyridine)(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dihydrate, and isothiocyanate (FITC).

[0039] In this embodiment, the fluorescence emission of the selected reference signal material is unaffected by the presence or concentration of wormwood pollen allergen in the detection system, providing a stable reference signal during detection. The fluorescence emission wavelength of the reference signal material differs significantly from that of the functionalized carbon quantum dots, forming distinguishable dual fluorescence emission peaks. This wavelength differentiation allows for quantitative analysis of wormwood pollen allergens by calculating the ratio of the two fluorescence peak intensities, avoiding errors caused by absolute intensity variations in single fluorescence signal detection and significantly improving detection sensitivity and accuracy. Furthermore, the aforementioned reference signal material can stably coexist with the response signal material (carbon quantum dots) in solution, thus forming a stable binding system with carbon quantum dots during the preparation of the proportional fluorescent probe.

[0040] As a further preferred design of this embodiment, the carrier includes one or more of nonwoven fabric, spunlace fabric, filter paper, nitrocellulose membrane, and polyvinylidene fluoride membrane. In this embodiment, one of the functions of the carrier is to enable long-term storage of the sensor. The carrier selected in this embodiment can fully retain the proportional fluorescence probe and has good corrosion resistance.

[0041] Correspondingly, according to a second aspect of the invention, referring to Figure 1 As shown, Figure 1 This is a flowchart illustrating the steps of preparing the allergen detection sensor according to the present invention. Figure 1 As shown, the present invention also provides a method for preparing an allergen detection sensor to prepare the allergen detection sensor provided in the first aspect of the present invention. The method includes the following steps: S1. The response signal material is prepared into a first solution of a certain concentration by adding ultrapure water; wherein, the response signal material is a carbon quantum dot material prepared based on the plant Xanthium sibiricum, and the carbon quantum dot material has the ability to specifically recognize the target allergen.

[0042] S2. Dissolve the reference signal material in a solvent to prepare a second solution of a certain concentration. The reference signal material includes one or more of europium trichloride hexahydrate, rhodamine B, platinum porphyrin, tridichloro-tris(2,2'-bipyridine)ruthenium(II) hexahydrate, dichloro-tris(1,10-phenanthroline)ruthenium(II) tetrahydrate, dichloro-bis(2,2'-bipyridine)(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dihydrate, and isothiocyanate (FITC).

[0043] S3. Under a nitrogen atmosphere, the first solution and the second solution are mixed in a certain proportion and stirred at room temperature for a certain time to combine the carbon quantum dot material and the response signal material.

[0044] S4. After the reaction is complete, transfer the reaction system to a dialysis bag and dialyze with excess ultrapure water for a certain period of time to obtain a proportional fluorescent probe.

[0045] S5. Load the proportional fluorescent probe onto the carrier to prepare the allergen detection sensor.

[0046] In this embodiment, the preparation method for the allergen detection sensor is simple, acts rapidly on the target allergen (such as wormwood pollen), has a specificity of over 95%, and provides results in 3 minutes.

[0047] Furthermore, step S4 includes: dialysis time of 1 day to 4 days.

[0048] Further, step S5 includes: loading the proportional fluorescent probe onto the carrier using one or more of the following methods: impregnation, electrostatic binding, and adsorption. The carrier includes one or more of the following: nonwoven fabric, spunlace cloth, filter paper, nitrocellulose membrane, and polyvinylidene fluoride membrane. A suitable loading method is fundamental to ensuring the activity of the proportional fluorescent probe. Improper loading methods may damage the probe's molecular structure and active groups, leading to a loss of specific recognition and fluorescence response to wormwood pollen allergens, thus rendering the sensor ineffective. The selected loading method maximizes probe stability, ensuring the practical application of the sensor.

[0049] Further, step S5 also includes: loading the proportional fluorescent probe onto the support for a loading time of 1 h to 5 h and a loading temperature of 5 °C to 45 °C. The loading time affects the degree of probe loading. If the loading time is too short, the interaction between the probe and the support will not be sufficient, resulting in insufficient loading and weak binding. This can easily lead to probe detachment during subsequent detection or storage, causing signal fluctuations and affecting the accuracy of the detection results. Conversely, if the loading time is too long, it may cause excessive reaction, leading to probe molecule aggregation on the support surface. A moderate loading time avoids aggregation and ensures sufficient loading. The loading temperature affects the probe's loading activity. Excessively high temperatures may damage the structural integrity of the reference and response signal materials in the proportional fluorescent probe, leading to an imbalance in their interaction. Insufficient temperatures will slow down the interaction rate between the probe and the support, preventing the probe from binding sufficiently and firmly to the support. The selected loading temperature ensures sufficient probe loading on the selected support and maintains its activity.

[0050] Further, step S1 includes: the concentration of the carbon quantum dot solution is preferably 0.001 mg / mL to 3 mg / mL. The selected concentration of carbon quantum dots is moderate, which will not cause aggregation due to excessive concentration, thereby reducing the sensing effect, and can maximize the rapid sensing of Artemisia pollen allergens.

[0051] Furthermore, step S2 includes using solvents such as methanol, acetonitrile, ultrapure water, and ethanol. The selected solvents exhibit good solubility for the reference fluorescent material, maximizing material dispersion.

[0052] Further, step S3 includes: the ratio of the first solution to the second solution is preferably 10:1 to 1:10. In proportional fluorescent probes, the ratio of the two materials has a significant impact on probe performance. A suitable ratio can form dual emission peaks and subsequently induce changes in probe color across different color systems.

[0053] Furthermore, step S3 also includes: a stirring time of 1 hour to 5 hours, and a stirring speed preferably of 100 rpm to 500 rpm. Both stirring speed and stirring time affect the bonding of the two materials. The selected stirring speed and stirring time are appropriate, which can ensure that the two materials are fully mixed.

[0054] It should be noted that, for the method embodiments, the embodiments of this application are not limited to the described order of actions, because according to the embodiments of this application, some steps may be performed in other orders or simultaneously.

[0055] According to another aspect of this application, such as Figure 5 and Figure 6 As shown, Figure 5 A schematic diagram of the structure of the rapid allergen detection device of the present invention is shown; Figure 6 A schematic diagram illustrating the method of using the rapid allergen detection device of the present invention is shown. The present invention also provides a rapid allergen detection device, comprising: an allergen detection sensor, such as the allergen detection sensor provided in the first aspect of the present invention; a dark box 2 with a notch in the side wall and a light-transmitting window 5 at the bottom; a support body 3, which enters the dark box 2 through the notch and is sealed and snapped onto the notch wall of the dark box 2; the support body 3 has a slot for holding the allergen detection sensor; a light source 1, disposed on the top wall of the dark box 2; and a camera device disposed below the bottom wall of the dark box 2; wherein the lens of the camera device, the light source 1, the light-transmitting window 5, and the allergen detection sensor are positioned opposite each other.

[0056] In this embodiment, the device consists of an allergen detection sensor, a dark box 2, a light source 1, and a camera device (such as a mobile phone and a mobile phone holder 4). The dark box 2 is a rectangular sealed structure made of light-absorbing material. A circular light-transmitting window 5 is opened at the bottom, and a notch can be opened in the middle of the side wall, into which a support body 3 is inserted. The support body 3 is long and narrow, with one end curved to form a circular slot, in which the allergen detection sensor can be placed. When the notch is inserted into the support body 3, the support body 3 is completely located inside the dark box 2, and the circular slot at the end is in the middle area of ​​the dark box 2. The other end of the notch, away from the slot, fits into the notch, sealing the dark box 2 again to create a closed, light-proof environment and prevent interference from stray light from the outside on the fluorescence signal. The support body 3 and the dark box 2 are designed as a pluggable structure, which facilitates the replacement of the sensor.

[0057] The slot houses the allergen detection sensor, which is the core detection element of the device. It is made based on carbon quantum dot-based proportional fluorescent probe technology that is specific to Artemisia pollen. It can specifically bind to Artemisia pollen allergens in the sample and trigger changes in fluorescence signals, which directly determines the specificity and sensitivity of the detection. It is the key to achieving allergen identification and signal response.

[0058] Light source 1 is a UV-LED lamp, preferably with a wavelength range of 200nm to 390nm. Light source 1 is used to provide specific wavelength light to excite the proportional fluorescent probe to generate a fluorescent signal. The fluorescent probe needs to be excited by the specific light source 1 to emit detectable fluorescence. The stability and wavelength matching of light source 1 directly affect the intensity and stability of the fluorescence signal, and are the energy source that ensures the probe can generate a signal normally.

[0059] The card slot located in the central area of ​​the dark box 2, the allergen detection sensor inside the card slot, the light-transmitting window 5 at the bottom of the dark box 2, and the light source 1 at the top of the dark box 2 are arranged coaxially.

[0060] In this embodiment, an external camera device is placed directly below the light-transmitting window 5. This camera device can be an industrial camera or a home video camera. The lens is aimed at the light-transmitting window 5, allowing light to pass through and directly reach the lens, facilitating the acquisition of images from the allergen detection sensor. After acquiring the images, the RGB intensity ratio is extracted using image analysis software, enabling further color quantification. Combined with a standard curve, the concentration of allergens can be further quantified. Therefore, this device ensures image acquisition accuracy, and by calculating fluorescence intensity ratios or color changes, quantitative and visual detection of allergens can be achieved.

[0061] Preferably, the camera device includes a mobile phone holder 4 and a mobile phone. The mobile phone holder 4 is positioned below the dark box 2, and the SIM card is attached to the mobile phone holder 4. The mobile phone has a built-in lens and an image processing unit. The lens is used to capture the fluorescence image generated when the allergen detection sensor comes into contact with the target allergen under the excitation of the light source 1. The image processing unit is used to convert the colors in the fluorescence image into RGB values ​​so as to obtain the concentration of the target allergen by combining it with a standard curve. The standard curve represents the trend of the RGB values ​​of the target allergen at different concentrations.

[0062] In this embodiment, the phone holder 4 has a double-slot structure. The two edges of the smartphone are inserted into the phone holder 4 along the slots on both sides, ensuring that the phone's lens is precisely aligned with the sensor detection area inside the dark box 2. This guarantees that the relative position of the phone and the sensor is consistent during each detection, thereby stabilizing the acquisition of fluorescence images. The phone has a built-in image processing unit that can quickly read the sensor's color, convert the color into RGB values, and then combine it with a standard curve to find the concentration of the corresponding allergen through the RGB values, thereby achieving rapid calculation of the allergen concentration.

[0063] As we know, the standard curve is a curve obtained by the inventors through numerous experiments, measuring the relationship between the concentration of the target allergen and the color parameters of the fluorescence image (such as RGB values ​​or the R / (B+G) ratio). Then, during actual testing, the RGB value of the current sample is read and compared with the standard curve to quickly calculate the actual concentration of the allergen in the current sample.

[0064] Rapid detection technology based on smartphones provides a convenient and efficient approach to detecting wormwood pollen allergens. When the sensor specifically binds to the wormwood pollen allergen in the sample, a proportional fluorescent probe produces an observable change in fluorescence signal, which is captured by the smartphone's camera. Using the phone's built-in image sensor and color recognition software, this color change is digitized into RGB values, and combined with a standard curve, the allergen concentration can be quickly detected. The entire process requires no complex operation and can be easily completed by ordinary users. This technology not only eliminates the reliance on large instruments in traditional detection methods but also enables on-site testing thanks to the portability of smartphones.

[0065] It should be noted that, for the above-described method and apparatus embodiments, since the allergen detection sensor already possesses the technical effects mentioned above, the preparation method for obtaining the allergen detection sensor and the apparatus containing the allergen detection sensor should also possess similar technical effects, so they will not be described in detail here.

[0066] The technical solution of the present invention will be described in more detail below with reference to several embodiments and accompanying drawings. It should be noted that, unless otherwise specified, the raw materials, chemical reagents and equipment used in the following embodiments can be obtained through market purchases, and the characterization methods can be implemented in accordance with methods known in the art.

[0067] Example 1 A method for preparing an allergen detection sensor, wherein the allergen detection sensor is prepared by combining carbon quantum dots with proportional fluorescent probes that are specific to Artemisia pollen, and then loading them onto a specific carrier. Specifically, the preparation method includes the following steps: S101. Prepare a first solution of 0.013 mg / mL by adding ultrapure water to the carbon quantum dot solution; S102. Dissolve europium trichloride hexahydrate in ethanol to prepare a second solution with a concentration of 0.2 mg / mL; S103. Under a nitrogen atmosphere, the first solution and the second solution are mixed in a ratio of 1:5 and stirred at room temperature for 2 hours at a stirring speed of 350 rpm. S104. After the reaction is complete, the reaction system is transferred to a dialysis bag and dialyzed with excess ultrapure water for 4 days to obtain a proportional fluorescent probe. S105. A novel wormwood allergen detection sensor was obtained by loading a proportional fluorescent probe onto a nonwoven fabric using an electrostatic bonding method at a loading temperature of 16℃ and a loading time of 3h.

[0068] The successful synthesis of proportional fluorescent probes is key to the preparation of sensors. Figure 2 The proportional fluorescent probe prepared in Example 1 and the fluorescence spectra of the two raw materials are shown. Figure 2 As shown, the characteristic peak of the carbon quantum dot is the emission peak at 430 nm, the reference signal is the emission peak at 600 nm, and the proportional fluorescent probe exhibits dual emission peaks at 430 nm and 600 nm under the excitation of a 300 nm ultraviolet lamp, proving its successful synthesis.

[0069] Performance testing: The proportional fluorescent probe prepared in Example 1 was used to detect the response time of Artv1 protein solution: Deionized water, Artv1 protein solution, buffer solution, and a proportional fluorescent probe solution were added to centrifuge tubes. The mixture was vortexed at a controlled temperature and shaking speed for different times (0.5 min, 1 min, 2 min, 3 min, 4 min, 5 min, 10 min, 20 min, 30 min, 60 min), and the fluorescence intensity changes of the probe were detected using a fluorescence spectrometer to determine the response time. Results are as follows: Figure 3 As shown, it can be seen that the Artv1 protein responds rapidly to the probe within the first 5 minutes, and the results are obtained after 5 minutes.

[0070] The proportional fluorescent probe prepared in Example 1 was used for the specific response detection of different protein solutions: Deionized water, different protein solutions (Artv1 protein solution, immunoglobulin A (IgA), hemoglobin (HGB), β-globulin, peanut allergen (Arah1)), buffer solution, and proportional fluorescent probe solution were added to centrifuge tubes. The mixture was shaken for 60 min at a controlled temperature and shaking speed, and the intensity changes of the proportional fluorescent probe were detected using fluorescence spectroscopy. The results are as follows: Figure 4 As shown, the response of immunoglobulin A (IgA), hemoglobin (HGB), β-globulin, and Arah1 to the proportional fluorescent probe in plasma was extremely low, less than 5%, while the response to Artv1 was 95%. This indicates that the wormwood pollen allergy sensor is specific to Artv1 and can specifically detect wormwood pollen allergens.

[0071] Example 2 A method for preparing an allergen detection sensor includes the following steps: S201. Prepare a first solution of 0.59 mg / mL by adding ultrapure water to the carbon quantum dot solution; S202, Dissolve isothiocyanate (FITC) in methanol to prepare a second solution with a concentration of 0.05 mg / mL; S203. Under a nitrogen atmosphere, the first solution and the second solution are mixed in a ratio of 5:2 and stirred at room temperature for 4 hours at a stirring speed of 250 rpm. S204. After the reaction is complete, the reaction system is transferred to a dialysis bag and dialyzed with excess ultrapure water for 6 days to obtain a proportional fluorescent probe. S205. A novel wormwood allergen detection sensor was obtained by loading a proportional fluorescent probe onto PVDF using an electrostatic bonding method at a loading temperature of 25℃ and a loading time of 4h.

[0072] The novel wormwood allergen detection sensor prepared in this embodiment has a specificity of 85% for the wormwood pollen allergen Artv1.

[0073] Example 3 A method for preparing an allergen detection sensor includes the following steps: S301. Prepare a first solution of 1 mg / mL by adding ultrapure water to the carbon quantum dot solution; S302. Dissolve tridichloro-tris(2,2'-bipyridine)ruthenium(II) hexahydrate in acetonitrile to prepare a second solution of 0.5 mg / mL; S303. Under a nitrogen atmosphere, the first solution and the second solution are mixed in a ratio of 9:2 and stirred at room temperature for 3 hours at a stirring speed of 320 rpm. S304. After the reaction is complete, the reaction system is transferred to a dialysis bag and dialyzed with excess ultrapure water for 1 day to obtain a proportional fluorescent probe. S305. A novel wormwood allergen detection sensor was obtained by loading a proportional fluorescent probe onto filter paper using an adsorption method at a loading temperature of 19℃ and a loading time of 2h.

[0074] The novel wormwood allergen detection sensor prepared in this embodiment has a specificity of 89% for the wormwood pollen allergen Artv1.

[0075] Example 4 A method for preparing an allergen detection sensor includes the following steps: S401. Prepare a first solution of 2 mg / mL by adding ultrapure water to the carbon quantum dot solution. S402. Dissolve Rhodamine B in ethanol to prepare a second solution with a concentration of 0.02 mg / mL; S403. Under a nitrogen atmosphere, the first solution and the second solution are mixed in a ratio of 2:7 and stirred at room temperature for 3 hours at a stirring speed of 220 rpm. S404. After the reaction is complete, the reaction system is transferred to a dialysis bag and dialyzed with excess ultrapure water for 3 days to obtain a proportional fluorescent probe. S405. A novel wormwood allergen detection sensor was obtained by loading a proportional fluorescent probe onto a nitrocellulose membrane using an impregnation method at a loading temperature of 35℃ and a loading time of 10h.

[0076] The novel wormwood allergen detection sensor prepared in this embodiment has a specificity of 90% for the wormwood pollen allergen Artv1.

[0077] Example 5 For instructions on using a rapid detection device for wormwood pollen allergens, please refer again. Figure 5 and Figure 6 The method of use is as follows: Dilute the body fluid of the allergic person with buffer solution, drop it onto the new allergen sensor, and then place it at the place where the new sensor is placed. Then place the mobile phone with the color recognition software application installed on the mobile phone holder 4, turn on the light source 1, take an image with the mobile phone and identify the color as RGB value, and then determine whether there is an allergy to wormwood pollen and calculate the allergen concentration based on the color change.

[0078] Example 6 A detection method for a rapid pollen allergen detection device is disclosed, which uses the device to identify and detect allergens at different concentrations. Figure 7 As shown, within the concentration range of 0 mM to 80 mM, the RGB values ​​of the detection parameters exhibit a good linear relationship with the allergen concentration. 2 A value of 0.999 indicates the reliability of the allergen sensor, making it suitable for use as a novel allergen detection device.

[0079] In summary, the wormwood pollen allergy sensor provided in this embodiment has the advantages of simple preparation, inexpensive and readily available raw materials, rapid action, good specificity, and visual detection. It can be used to detect the wormwood pollen allergen protein Artv1, showing promise as a novel allergy detection technology. It can solve the problems of long detection time, high cost, and low accuracy in current wormwood pollen allergy detection methods. The rapid detection device, combined with a smartphone camera, quickly reads the sensor's color and uses mobile phone color recognition software to read the RGB values ​​of the color. Based on the provided reference data, it quickly calculates the allergen concentration, offering convenience and speed, and showing promise for clinical application.

[0080] It should also be noted that, in this document, the terms "upper," "lower," "left," "right," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing the invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of the invention. Furthermore, relational terms such as "first" and "second" are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations, nor should they be construed as indicating or implying relative importance. Moreover, the term "comprising" or any other variation thereof is intended to cover non-exclusive inclusion, such that a process, method, article, or terminal device that comprises a list of elements includes not only those elements, but also other elements not expressly listed, or elements inherent to such a process, method, article, or terminal device.

Claims

1. An allergen detection sensor, characterized in that, The sensors include: The response signal material is a carbon quantum dot material prepared based on the plant Xanthium sibiricum, and the carbon quantum dot material has the ability to specifically recognize the target allergen. A reference signal material, wherein the reference signal material is combined with the carbon quantum dot material to form a proportional fluorescent probe; The carrier on which the proportional fluorescent probe is loaded.

2. The allergen detection sensor according to claim 1, characterized in that, The reference signal material includes one or more of europium trichloride hexahydrate, rhodamine B, platinum porphyrin, tridichloro-tris(2,2'-bipyridine)ruthenium(II) hexahydrate, dichloro-tris(1,10-phenanthroline)ruthenium(II) tetrahydrate, dichloro-bis(2,2'-bipyridine)(4,7-diphenyl-1,10-phenanthroline)ruthenium(II) dihydrate, and isothiocyanate (FITC).

3. The allergen detection sensor according to claim 1, characterized in that, The carrier includes one or more of the following: nonwoven fabric, spun cloth, filter paper, nitrocellulose membrane, and polyvinylidene fluoride membrane.

4. A method for preparing an allergen detection sensor, characterized in that, Preparation methods include: The response signal material is prepared into a first solution of a certain concentration by adding ultrapure water; wherein, the response signal material is a carbon quantum dot material prepared based on the plant Xanthium sibiricum, and the carbon quantum dot material has the ability to specifically recognize the target allergen; The reference signal material is dissolved in a solvent to prepare a second solution of a certain concentration; Under a nitrogen atmosphere, the first solution and the second solution are mixed in a certain proportion and stirred at room temperature for a certain time to combine the carbon quantum dot material and the response signal material. After the reaction was completed, the reaction system was transferred to a dialysis bag and dialyzed with excess ultrapure water for a certain period of time to obtain a proportional fluorescent probe. The proportional fluorescent probe is loaded onto a carrier to prepare an allergen detection sensor.

5. The method for preparing an allergen detection sensor according to claim 4, characterized in that, Loading methods for loading the proportional fluorescent probe onto the carrier include one or more of the following: impregnation, electrostatic binding, and adsorption.

6. The method for preparing an allergen detection sensor according to claim 4, characterized in that, The solvent includes any one of methanol, acetonitrile, ultrapure water, and ethanol.

7. The method for preparing an allergen detection sensor according to claim 4, characterized in that, The concentration of the first solution is 0.001 mg / mL to 3 mg / mL; the ratio of the first solution to the second solution is 10:1 to 1:10; the stirring time is 1 h to 5 h, and the stirring speed is 100 rpm to 500 rpm.

8. The method for preparing an allergen detection sensor according to claim 4, characterized in that, During the process of loading the proportional fluorescent probe onto the carrier, the loading time is 1h to 5h and the loading temperature is 5℃ to 45℃.

9. A rapid allergen detection device, characterized in that, The device includes: An allergen detection sensor, as described in any one of claims 1-3; The dark box has notches in the side walls and a light-transmitting window at the bottom. The support body enters the dark box through the notch and is sealed and snapped onto the notch wall of the dark box; the support body has a slot in which the allergen detection sensor is placed. The light source is located on the top wall of the darkroom; The camera device is located below the bottom wall of the darkroom; The camera lens, the light source, the light-transmitting window, and the allergen detection sensor are positioned relative to each other.

10. The rapid allergen detection device according to claim 9, characterized in that, The camera device includes a mobile phone holder and a mobile phone. The mobile phone holder is located below the dark box, and the SIM card is connected to the mobile phone holder. The mobile phone has a built-in camera and image processing unit; wherein... The lens is used to capture the fluorescence image generated by the allergen detection sensor when it comes into contact with the target allergen under the excitation of the light source; The image processing unit is used to convert the colors in the fluorescence image into RGB values ​​so as to obtain the concentration of the target allergen by combining it with a standard curve; the standard curve represents the trend of the RGB values ​​of the target allergen at different concentrations.

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