Optical disc-based method for quantitative detection of biological samples

Abstract

The application discloses a bio-sample quantitative detection method based on an optical disc and belongs to the technical field of biosensors. The method comprises the following steps: for a mixed system to-be-detected solution, based on the inhibition or promotion effect of the solution on an insoluble precipitate solution, the solution is reacted with the insoluble precipitate, and a suspension of the remaining insoluble precipitate after the reaction is prepared; after the surface of an optical disc full of data is subjected to an activation treatment, the insoluble precipitate suspension corresponding to the mixed system to-be-detected solution or a single system to-be-detected solution is added dropwise on the optical disc surface at a specified radius to form a sample drop point; after the water in the sample drop point is completely evaporated, the data in the optical disc is read, data reading errors are obtained, and corresponding LDC counts are calculated; and the LDC counts are substituted into a pre-calibrated standard curve or a relational expression to obtain the concentration of the to-be-detected solution. The application can improve the efficiency and universality of quantitative detection of substances in a solution and reduce detection cost and detection complexity.

Description

Optical disc-based quantitative detection method for biological samples Technical Field

[0001] This invention belongs to the field of biosensor technology, and more specifically, relates to a method for quantitative detection of biological samples based on optical discs. Background Technology

[0002] Currently, biochemical analysis has become an important task in the field of analytical science. Existing methods such as mass spectrometry, spectral analysis, chromatography, and enzyme-linked immunosorbent assay (ELISA) have made significant progress in accuracy, sensitivity, throughput, and automation. However, traditional biochemical analysis techniques are limited by expensive instruments and specialized operations, mainly confining them to laboratories in developed regions. Adaptive use strategies, which utilize the physical mechanisms of everyday items, can reduce testing costs, simplify operations, and improve accessibility and versatility. This is of great significance for early field biomedical analysis, especially in resource-limited areas.

[0003] In recent years, consumer electronics products such as optical drives have been adapted into tools for point-of-care testing (POCT), particularly in the field of biochemical assays. The sophisticated optics and servo systems of optical drives enable optical discs to serve as substrates for biochemical reactions, leading to the development of various optical disc-based biochemical assay strategies. While these strategies are innovative, they often require customization or modification of the optical drive, optical disc, or software, which limits their widespread application to some extent. Furthermore, complex quantitative dispensing operations, high barriers to entry, insufficient integration, high reagent consumption, slow detection speed, and high costs remain challenges for the technology.

[0004] To address the aforementioned issues, researchers have proposed a strategy that does not require hardware modifications or new software development. However, the process of constructing a reaction system on the surface of Blu-ray Disc (BD) remains cumbersome and time-consuming, requiring multiple reaction steps (such as surface activation, probe immobilization, sample addition and sealing) and additional labeling reagents (such as silver staining). This not only increases operational errors but also raises the professional requirements for operators, thus limiting the promotion and application of these technologies to some extent. Summary of the Invention

[0005] In response to the shortcomings and improvement needs of existing technologies, this invention provides a method for quantitative detection of biological samples based on optical discs. The purpose is to improve the efficiency and versatility of quantitative detection of substances in solution, and reduce detection costs and complexity.

[0006] To achieve the above objectives, according to one aspect of the present invention, a method for quantitative detection of biological samples based on optical discs is provided for detecting the concentration of a test solution; the test solution is a soluble target solution or an insoluble precipitate suspension, and neither the soluble target in the soluble target solution nor the insoluble precipitate in the insoluble precipitate suspension will damage the surface of the optical disc; the optical disc is filled with data; the quantitative detection method includes:

[0007] After the optical disc is activated, the test solution is dropped onto a specified radius on the surface of the optical disc to form a sample dropping point; the activation treatment changes the surface of the optical disc from hydrophobic to hydrophilic.

[0008] After the water in the sample droplet has completely evaporated, the data in the optical disc is read, the data read errors are obtained, and the integral area of ​​the signal peak of the data read error is calculated to obtain the corresponding LDC count.

[0009] The concentration corresponding to the calculated LDC count is determined according to the pre-calibrated standard curve, and the concentration of the test solution is obtained.

[0010] When the test solution is a soluble target solution, the standard curve is obtained by fitting the LDC counts corresponding to different concentrations of soluble target solutions; when the test solution is an insoluble precipitate suspension, the standard curve is obtained by fitting the LDC counts corresponding to different concentrations of insoluble precipitate suspensions.

[0011] Furthermore, the calibration methods for standard curves include:

[0012] Prepare test solutions with different concentrations as standard solutions;

[0013] After the optical disc is activated, the standard solution is uniformly added to the surface of the optical disc along a specified radius to obtain the sample dropping point corresponding to each standard solution;

[0014] After all the water in the sample drop points has completely evaporated, the data in the optical disc is read, the data reading errors corresponding to each sample drop point are obtained, and the integral area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution.

[0015] By fitting the concentrations of each standard solution with their corresponding LDC counts, a standard curve is obtained to record the relationship between concentrations and LDC counts.

[0016] According to another aspect of the present invention, a method for quantitative detection of biological samples based on optical discs is provided, for detecting the concentration of a test solution, wherein the test solution is a soluble target solution; the optical disc is filled with data; the quantitative detection method includes:

[0017] Step S1: Mix the test solution with an excess of insoluble precipitate suspension to allow the soluble target in the test solution to react fully with the insoluble precipitate in the insoluble precipitate suspension; the soluble target can react with the insoluble precipitate, and a soluble product is produced after the reaction; the insoluble precipitate will not damage the surface of the optical disc;

[0018] Step S2: Centrifuge the mixed solution;

[0019] Step S3: After removing the supernatant from the centrifuged mixed solution, add deionized water to the remaining solution to obtain a new suspension of insoluble precipitate;

[0020] Step S4: After the optical disc is activated, the insoluble precipitate suspension obtained in step S3 is dropped onto the optical disc surface at a specified radius to form sample dropping points; the activation treatment changes the surface of the optical disc from hydrophobic to hydrophilic.

[0021] Step S5: After the water in the sample droplets has completely evaporated, read the data from the optical disc, obtain the data reading errors, and calculate the integral area of ​​the signal peak of the data reading error to obtain the corresponding LDC count;

[0022] Step S6: Substitute the LDC count obtained in step S5 into the pre-calibrated relationship between the LDC count and the concentration of the solution to be tested to obtain the concentration of the solution to be tested.

[0023] Furthermore, the calibration methods for the relationship between LDC count and the concentration of the solution to be tested include:

[0024] Prepare test solutions with different concentrations as standard solutions;

[0025] Each standard solution was mixed separately with an excess of an insoluble precipitate suspension of equal concentration to ensure that the soluble target in each standard solution reacted fully with the insoluble precipitate in the insoluble precipitate suspension.

[0026] Each mixed solution was centrifuged separately.

[0027] After removing the supernatant from the centrifuged mixed solution, deionized water was added to the remaining solution to obtain new suspensions of insoluble precipitates corresponding to each standard solution.

[0028] After the optical disc is activated, the insoluble precipitate suspensions corresponding to each standard solution are uniformly added to the surface of the optical disc along a specified radius to obtain the sample dropping points corresponding to each standard solution.

[0029] After all the water in the sample drop points has completely evaporated, the data in the optical disc is read, the data reading errors corresponding to each sample drop point are obtained, and the integral area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution.

[0030] By fitting the concentrations of each standard solution with the corresponding LDC counts, the relationship between LDC counts and the concentrations of the solution to be tested is obtained.

[0031] According to another aspect of the present invention, a method for quantitative detection of biological samples based on optical discs is provided, for detecting the concentration of a test solution, wherein the test solution is a soluble target solution; the optical disc is filled with data; the quantitative detection method includes:

[0032] Step T1: Mix the test solution, the insoluble precipitate suspension, and the third reactant solution to ensure that the insoluble precipitate in the insoluble precipitate suspension reacts fully with the third reactant in the third reactant solution; the insoluble precipitate reacts with the third reactant, producing a soluble product after the reaction; the insoluble precipitate suspension is in excess relative to the third reactant solution; the test solution inhibits the reaction between the third reactant and the insoluble precipitate, and the higher the concentration of the test solution, the stronger the inhibitory effect; the insoluble precipitate will not damage the surface of the optical disc;

[0033] Step T2: Centrifuge the mixed solution;

[0034] Step T3: After removing the supernatant from the centrifuged mixed solution, add deionized water to the remaining solution to obtain a new suspension of insoluble precipitate;

[0035] Step T4: After the optical disc is activated, the insoluble precipitate suspension obtained in step T3 is dropped onto the optical disc surface at a specified radius to form sample drop points; the activation treatment changes the surface of the optical disc from hydrophobic to hydrophilic.

[0036] Step T5: After the water in the sample dropper has completely evaporated, read the data from the optical disc, obtain the data read errors, and calculate the integral area of ​​the signal peak of the data read error to obtain the corresponding LDC count;

[0037] Step T6: Substitute the LDC count obtained in step T5 into the pre-calibrated relationship between the LDC count and the concentration of the solution to be tested to obtain the concentration of the solution to be tested.

[0038] Furthermore, the calibration methods for the relationship between LDC count and the concentration of the solution to be tested include:

[0039] Prepare test solutions with different concentrations as standard solutions;

[0040] For each standard solution, it is mixed with the insoluble precipitate suspension and the third reactant solution to ensure that the insoluble precipitate reacts fully with the third reactant; the concentration of the insoluble precipitate suspension mixed with each standard solution is equal, and the concentration of the third reactant solution mixed with each standard solution is equal.

[0041] Each mixed solution was centrifuged separately.

[0042] After removing the supernatant from the centrifuged mixed solution, deionized water was added to the remaining solution to obtain new suspensions of insoluble precipitates corresponding to each standard solution.

[0043] After the optical disc is activated, the insoluble precipitate suspensions corresponding to each standard solution are uniformly added to the surface of the optical disc along a specified radius to obtain the sample dropping points corresponding to each standard solution.

[0044] After all the water in the sample drop points has completely evaporated, the data in the optical disc is read, the data reading errors corresponding to each sample drop point are obtained, and the integral area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution.

[0045] By fitting the concentrations of each standard solution with the corresponding LDC counts, the relationship between LDC counts and the concentrations of the solution to be tested is obtained.

[0046] According to another aspect of the present invention, a method for quantitative detection of biological samples based on optical discs is provided for detecting the concentration of bacterial solutions; the optical discs are filled with data; the quantitative detection method includes:

[0047] After the optical disc is activated, a bacterial solution is dropped onto the surface of the disc at a specified radius to form sample droplets. The activation treatment changes the surface of the optical disc from hydrophobic to hydrophilic. After activation, the ability of the optical disc surface to adsorb bacteria is enhanced.

[0048] After the water in the sample droplet has completely evaporated, the data in the optical disc is read, the data read errors are obtained, and the integral area of ​​the signal peak of the data read error is calculated to obtain the corresponding LDC count.

[0049] The concentration of the bacterial solution is obtained by determining the concentration corresponding to the calculated LDC count according to the pre-calibrated standard curve.

[0050] The standard curve was obtained by fitting the LDC counts corresponding to bacterial solutions of different concentrations.

[0051] Furthermore, the calibration methods for standard curves include:

[0052] Bacterial solutions of different concentrations were prepared as standard solutions;

[0053] After the optical disc is activated, the standard solution is uniformly added to the surface of the optical disc along a specified radius to obtain the sample dropping point corresponding to each standard solution;

[0054] After all the water in the sample drop points has completely evaporated, the data in the optical disc is read, the data reading errors corresponding to each sample drop point are obtained, and the integral area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution.

[0055] By fitting the concentrations of each standard solution with their corresponding LDC counts, a standard curve is obtained to record the relationship between concentrations and LDC counts.

[0056] Furthermore, in the above-mentioned optical disc-based quantitative detection method for biological samples, the activation process includes:

[0057] The optical disc is immersed in NaOH solution for a preset time to generate reactive carboxyl groups on its surface, thus changing it from a hydrophobic to a hydrophilic state.

[0058] In summary, the technical solution conceived in this invention proposes a novel "drip-dry strategy" (2D), which involves directly adding a sample solution to the surface of an activated optical disc to form sample droplets. After the moisture in the sample droplets has completely evaporated, data is read, and quantitative detection results are obtained based on the data reading errors. The relevant detection principle is as follows: after the moisture in the sample droplets completely evaporates, the substances deposit on the optical disc surface to form a covering area, which prevents laser reflection and causes data reading errors when the optical drive reads the data. These data reading errors have a calibrable quantitative relationship with the sample concentration. Existing optical disc quality diagnostic software can be used to obtain the data reading errors caused by the sample droplets, after which quantitative detection can be completed.

[0059] Based on the proposed "direct spotting-drying" strategy, this invention employs a "direct spotting-drying" strategy for solutions that will not damage the surface of optical discs. This eliminates the need for customization or modification of the optical drive, optical disc, or software, as well as complex operations such as probe fixation and sealing. It also eliminates the need for additional labeling reagents such as silver staining. The operation is simple and convenient, with high detection efficiency, low detection complexity, and low cost. At the same time, there are no special operational requirements for the related operations, which greatly improves its versatility.

[0060] For complex mixed solutions or solutions that might damage the optical disc surface, an insoluble precipitate suspension that will not damage the disc surface is introduced. The reaction process of the insoluble precipitate is regulated by a soluble target in the test solution. Specifically, the soluble target reacts with the insoluble precipitate, or inhibits the reaction between the insoluble precipitate and other reactants, thus controlling the dissolution process and achieving effective purification of the insoluble precipitate. The concentration of the purified insoluble precipitate suspension is then detected indirectly through a "direct spotting-drying" strategy, enabling quantitative detection of the test solution. The entire purification process is performed outside the optical disc; only the "direct spotting-drying" strategy is applied to the disc surface. This method is simple and convenient, with high detection efficiency, low complexity, low cost, and high versatility.

[0061] Compared with the prior art, the present invention can achieve the following beneficial effects:

[0062] (1) When performing quantitative detection of biological samples, this invention only requires optical discs, standard optical drive equipment and optical disc quality diagnostic software. The relevant software and hardware are readily available, which greatly reduces the detection cost (cost per test < US$1).

[0063] (2) The entire detection process, including sample processing, error reading and result analysis, can be completed within 30 minutes. The sample processing operation is simple and easy to use, providing an alternative rapid substance analysis method for early, field and home use.

[0064] (3) This invention is universal and applicable to the quantification of almost all substances in a single system; in the case of a mixed system, this method is also applicable to most reactions that can produce or dissolve precipitates, transforming the quantification of the target substance into the quantification of separable and purifiable precipitates. This method can provide new ideas and approaches for substance analysis and research in many fields such as biomedicine, food hygiene, and environmental monitoring.

[0065] (4) The diameter of the laser spot on the surface of the optical disc can reach the micrometer level. For example, the Blu-ray optical disc uses a laser with a wavelength of 405nm. The data layer is only 0.1mm away from the surface, and the diameter of the laser spot on the surface is 138μm. Therefore, the biological sample quantitative detection method provided by the present invention can theoretically achieve a resolution of the micrometer level. High-throughput detection can be achieved by constructing more densely arranged detection sites on the surface of the optical disc.

[0066] In summary, this invention constructs a low-cost, high-efficiency quantitative detection system for biological samples by combining common optical drive equipment and free software. This system can complete the test within 30 minutes, is suitable for quantitative analysis of substances in multiple fields, and possesses high-throughput detection capabilities, providing a new approach for future research and applications. Attached Figure Description

[0067] Figure 1 is a schematic diagram of the software and hardware related to the quantitative detection method for biological samples provided by the present invention;

[0068] Figure 2 is a schematic diagram of the structure of the Blu-ray disc provided by the present invention and the basic principle of BD error reading; in the structure of each layer of the Blu-ray disc, 1 is the hard coating, 2 is the cover layer, 3 is the adhesive layer, 4 is the recording layer, 5 is the reflective layer, 6 is the substrate, and 7 is the protective layer.

[0069] Figure 3 is a schematic diagram of the process by which the optical disc quality diagnostic software used in this invention reads and analyzes BD errors, and then calculates the error rate.

[0070] Figure 4 is a schematic diagram of the operation process of the "direct spotting-drying" strategy (2D strategy) in this invention;

[0071] Figure 5 is a schematic diagram of the principle and operation process of the "direct spotting-drying" strategy provided by the present invention in three typical biomonitoring methods;

[0072] Figure 6 is a schematic diagram of the data results obtained by quantitative detection of bovine serum albumin provided in Example 1 of the present invention; wherein, (a) shows the original state, microstructure and LDC distribution of the BSA gradient solution when detecting BSA solution concentration using the traditional BCA kit method, (b) shows the original state, microstructure and LDC distribution of the BSA gradient solution when detecting BSA solution concentration using the "direct spotting-drying" strategy, (c) shows the dependence of LDC count and original solution gray value on protein concentration in the BCA kit method, and (d) shows the dependence of LDC count and original solution gray value on protein concentration under the "direct spotting-drying" strategy.

[0073] Figure 7 is a schematic diagram of the data results obtained by quantitative detection of pure MnO2 provided in Example 2 of the present invention; wherein, (a) is a photograph of MnO2 suspension, a super depth-of-field micrograph of sample drop points and LDC count distribution map, and (b) is a schematic diagram of the dependence of LDC value of sample drop points of MnO2 suspension and gray value of original solution on MnO2 concentration.

[0074] Figure 8 is a schematic diagram of the process and results of quantitative detection of GSH glutathione provided in Example 3 of the present invention; wherein, (a) is a schematic diagram of MnO2 purification, (b) is a schematic diagram of the reaction between GSH glutathione and MnO2, (c) is a TEM image of the gradient suspension of pure MnO2 and a schematic diagram of LDC distribution, (d) is a schematic diagram of the dependence between LDC count and GSH concentration, and (e) is a schematic diagram of the linear fitting between LDC count and GSH concentration.

[0075] Figure 9 is a schematic diagram illustrating the principle of quantitative detection of urea in human serum provided in Embodiment 4 of the present invention;

[0076] Figure 10 is a schematic diagram of the process and results of quantitative detection of organophosphorus pesticides provided in Example 5 of the present invention; wherein, (a) is a schematic diagram of the reaction between MnO2 and thioacetylcholine under the catalysis of acetylcholinesterase, (b) is a schematic diagram of the original state of the organophosphorus pesticide gradient solution and the corresponding LDC distribution of residual MnO2, (c) is a schematic diagram of the dependence between the concentration of organophosphorus pesticides (OPs) and the LDC count of residual MnO2, and (d) is a schematic diagram of the linear fitting results between the concentration of organophosphorus pesticides (OPs) and the LDC count of residual MnO2. Detailed Implementation

[0077] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention. Furthermore, the technical features involved in the various embodiments of this invention described below can be combined with each other as long as they do not conflict with each other.

[0078] It should be noted that in the description of this invention, the terms "upper," "lower," "left," "right," "inner," and "outer," etc., indicating directional or positional relationships, are based on the directional or positional relationships shown in the accompanying drawings. This is merely for ease of description and does not indicate or imply that the device or element must have a specific orientation, or be constructed and operated in a specific orientation; therefore, it should not be construed as a limitation of this invention. Furthermore, it should be noted that in the description of this invention, unless otherwise explicitly specified and limited, the terms "installed," "connected," and "linked" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal communication between two elements. The terms "first," "second," etc. (if present), in this invention and its accompanying drawings are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0079] To address the issues of complex operation and high cost of existing biochemical analysis techniques, this invention proposes a simple and efficient optical disc-based quantitative detection method for biological samples. This method, based on a standard optical disc and its drive, along with free optical disc quality diagnostic software, employs an innovative "drip-dry strategy" (2D) to provide an economical and practical solution for the rapid and quantitative detection of substances in solution. Specifically, the "drip-dry strategy" involves directly spotting the test solution onto the surface of an activated optical disc. As the water evaporates, substances in the solution form sample spots on the disc surface. These spots interfere with laser reflection and are thus identified as erroneous signals by the optical disc drive. Furthermore, a calibrable quantitative relationship exists between these erroneous signals and the solution concentration. Combined with optical disc quality diagnostic software, these erroneous signals can be accurately recorded and analyzed, thereby achieving accurate quantification of the substance content in the solution.

[0080] In practical applications, any type of optical disc, such as Blu-ray disc, CD, or DVD, along with its corresponding optical disc drive and optical disc quality diagnostic software, can be selected. Without loss of generality, the following embodiments will use Blu-ray discs as an example for illustration; correspondingly, the optical disc drive is a standard BD drive (Plextor BD), and the optical disc quality diagnostic software is PlexUTILITIES. The standard BD drive is responsible for reading and writing BD disc information, while the optical disc quality diagnostic software runs on a computer and is used to analyze the quality condition of the BD disc surface, especially data reading errors, as shown in Figure 1.

[0081] Compared to CD and DVD systems, Blu-ray drives use shorter wavelength (405nm) lasers and larger numerical aperture objectives, which continuously shrinks the laser focusing spot and increases storage density. In optical drive-based biomolecular detection, a smaller detection spot means higher detection sensitivity and resolution.

[0082] The Blu-ray disc, as the core of the information storage system, boasts a storage capacity of up to 23.31 GB. Its structure, as shown in Figure 2, is a multi-layered structure, consisting of, from top to bottom: a hard coating 1, a cover layer 2, an adhesive layer 3, a recording layer 4, a reflective layer 5, a substrate 6, and a protective layer 7. The data area within the disc is precisely located within a ring-shaped region with an outer radius of 58 mm and an inner radius of 24 mm. Utilizing sophisticated algorithms, the system can rapidly convert logical locations (in GB) into physical locations (in mm), achieving efficient data management. Furthermore, by employing Picket code error correction technology, the surface of the Blu-ray disc is meticulously divided into multiple error correction blocks of 64 kb each. These blocks are arranged in an orderly spiral pattern, ensuring that sample droplets can only be presented in a radial pattern.

[0083] BD drives use high-precision lasers to accurately record and read information from BD discs. During recording, the laser moves from the inside out along a spiral path on the disc surface, creating a series of fine pits on the data layer. These pits and the flat areas on the disc have significantly different reflectivity, which forms the basis of information encoding: when the reflectivity of a pit is the same as that of a flat area, it represents the binary number "0"; however, when the laser sweeps across the boundary between the pit and the flat area, the change in the direction and intensity of the reflected light represents the binary number "1".

[0084] The quality diagnostic software utilizes two testing modes—LDC (Long Distance Code) and BIS (Burst Indicator Code)—to monitor and quantify the number of errors within the error correction block. When the surface of a Blu-ray disc is affected by contaminants or sample droplets, the optical properties of the disc surface are significantly altered, causing interference to the laser beam during focusing, as shown in Figure 2. This interference phenomenon is visually represented on the software interface as LDC error count signal peaks. Each signal peak corresponds to a specific sample droplet, and the integral area of ​​the signal peaks directly reflects the LDC count caused by these droplets. The LDC count is the number of check bytes of the LDC codeword used for error correction, reflecting the magnitude of the error. This precise correspondence enables the present invention to achieve accurate and rapid detection of substances in solution. In practical applications, this correspondence can be calibrated by pre-preparing standard solutions.

[0085] In practical applications, this invention utilizes the burning function of the Windows system to burn a 22.3GB video file onto a blank Blu-ray disc, filling it with data. Then, it is immersed in a 0.1M NaOH solution for 1.5 hours and activated at 55°C, transforming the Blu-ray surface from hydrophobic to hydrophilic. This activation treatment, changing the Blu-ray surface from hydrophobic to hydrophilic, makes it easier for the sample droplets to form regular circles after drying, facilitating more accurate quantitative detection. Next, as shown in Figures 3 and 4, solutions of different concentrations (preferably with a concentration gradient) are prepared as standard solutions. If insoluble precipitates need to be introduced, they are introduced and purified. The treated standard solutions are then added radially along the Blu-ray surface to ensure uniform distribution on the hydrophilic surface. If insoluble precipitates are not required, the standard solutions are directly added radially along the Blu-ray surface to ensure uniform distribution. Subsequently, the sample is dried in a drying oven or at room temperature to ensure complete deposition of the substances in the solution onto the optical disc (BD), forming clearly identifiable sample droplets. Then, data reading errors are detected on the BD using optical disc quality diagnostic software. The signal peak integral area of ​​each error signal is calculated to obtain the LDC count corresponding to each standard solution. Finally, the concentration of the standard solution and the corresponding LDC count are fitted to obtain the corresponding curve or calculation expression, thus calibrating the corresponding quantitative relationship.

[0086] Based on the correspondence obtained from calibration, the solution to be tested or the treated solution is dropped onto the corresponding diameter of the BD surface to form sample drop points, following the same method. After the water in the sample drop points has completely evaporated, the data is read, data reading errors are obtained, and the LDC count is calculated. Substituting into the corresponding correspondence, the concentration of the solution to be tested can be obtained, thus achieving quantitative detection.

[0087] Based on the detection approach provided by this invention, as shown in Figure 5, for solutions with a single system that do not damage the surface of the optical disc, the "direct spotting-drying" strategy is directly applied for quantitative detection. For complex mixed system solutions or solutions that may damage the surface of the optical disc, an insoluble precipitate suspension that will not damage the optical disc surface is introduced. The reaction process of the insoluble precipitate is regulated by the soluble target in the test solution, that is, by using the soluble target to react with the insoluble precipitate, or by using the soluble target to inhibit the reaction between the insoluble precipitate and other reactants, thereby regulating the dissolution process of the insoluble precipitate and achieving effective purification of the precipitate. Then, the "direct spotting-drying" strategy is used to quantitatively detect the purified insoluble precipitate suspension, thereby indirectly achieving quantitative detection of the test solution.

[0088] This invention achieves rapid, accurate, and low-cost quantitative detection of substances in solution through a "direct spotting-drying" strategy. The entire detection process can be completed in just 30 minutes and requires no professional personnel, further reducing the cost and time of biomolecule detection. Furthermore, by using soluble targets to regulate the dissolution process of insoluble precipitates, it is also possible to achieve indirect quantitative detection of substances in complex mixed solutions, broadening its application scope.

[0089] The following detailed explanation is provided in conjunction with specific embodiments.

[0090] Example 1:

[0091] A method for quantitative detection of biological samples based on optical discs is used to achieve quantitative detection of bovine serum albumin (BSA) solution.

[0092] This embodiment also utilizes the traditional BCA kit method to quantitatively detect the same bovine serum albumin solution, and its detection process and results serve as a comparative example of this embodiment.

[0093] Figure 6(a) shows the initial solution state of the protein gradient solution with added BCA reagent when using the traditional BCA kit method. The microscopic morphology of the sample droplets was captured using a super depth-of-field microscopy image, which also shows the LDC distribution. Figure 6(c) visually illustrates the dependence of protein LDC count and initial solution grayscale value on protein concentration in the BCA method.

[0094] The photograph of the reaction solution shown in Figure 6(a) clearly demonstrates the significant color gradient changes formed by different concentrations of BSA. Grayscale analysis using ImageJ software revealed an acceptable linear relationship between grayscale values ​​and BSA concentration. However, after applying the drip-drying strategy, the LDC distribution remained roughly at the same level, as shown in Figure 6(c). Furthermore, the super-depth-of-field micrograph also showed that, despite slight pattern variations, the coverage of the samples at each point was almost insignificantly different. This observation is likely due to a crucial fact in the BCA colorimetric method: the concentration of the detection reagent is much higher than that of BSA. Therefore, during evaporation and drying, all the solutes (including the detection reagent and BSA) remain on the BD surface. This indicates that the drip-drying strategy does not rely on color changes as in traditional colorimetric methods, but rather on the total concentration of all substances in the solution.

[0095] Subsequently, in this embodiment, a pure BSA gradient solution was prepared, and the LDC count was calculated using the aforementioned "direct spotting-drying" strategy. Although the concentration differences of these solutions were visually difficult to distinguish, a clear gradient trend was observed in the LDC distribution of the sample drops, as shown in Figure 6(b). Calculations revealed a positive correlation between LDC count and BSA concentration, as shown in Figure 6(d). Although a saturation trend appeared at higher BSA concentrations, this still demonstrates the potential of the drip-drying strategy for quantitative detection of substances in a single system within a specific concentration range. It should be noted that this specific concentration range is sufficient to cover the concentration of the BSA solution actually required for measurement.

[0096] Using the curve obtained by fitting the BAS concentration and the corresponding LDC count shown in Figure 6(d) as the standard curve, this embodiment utilizes the "direct spotting-drying" strategy provided by the present invention for the quantitative detection method of bovine serum albumin, specifically including:

[0097] After the optical disc is activated, the bovine serum albumin solution to be tested is dropped onto a specified radius on the surface of the optical disc to form a sample dropping point; it is easy to understand that the diameter of the bovine serum albumin solution to be tested is the same as the diameter of the standard solution to be dropped when calibrating the standard curve;

[0098] After the water in the sample droplet has completely evaporated, the data in the optical disc is read, the data read errors are obtained, and the integral area of ​​the signal peak of the data read error is calculated to obtain the corresponding LDC count.

[0099] The concentration corresponding to the calculated LDC count is determined according to the pre-calibrated standard curve, and the concentration of the bovine serum albumin solution to be tested is obtained.

[0100] It should be noted that if the relevant standard curve has been calibrated in advance, there is no need to perform calibration again; the test can be performed directly based on the calibrated standard curve.

[0101] The comparison revealed that when using traditional BCA kits for protein quantification, proteins need to react with colorimetric reagents at room temperature for 1-2 hours before absorbance can be measured or colorimetric analysis performed. However, this embodiment completely eliminates this cumbersome reaction step, enabling direct quantification of pure proteins in approximately 20 minutes. This result fully demonstrates the significant potential of CD-ROM-based biological sample quantification methods assisted by the "drip-dry strategy" (2D) in rapid point-of-care testing (POCT).

[0102] Example 2:

[0103] A method for quantitative detection of biological samples based on optical discs is disclosed, used to achieve quantitative detection of MnO2 solution. This embodiment also employs the aforementioned "direct spotting-drying" strategy to directly quantify the MnO2 solution to be tested.

[0104] To calibrate a standard curve for recording the correlation between MnO2 solution concentration and LDC count, this embodiment used deionized water and MnO2 nanoparticles (particle size between 100 nm and 150 nm) to prepare eight suspensions with different concentration gradients in eight EP tubes (microcentrifuge tubes). The concentrations were 0, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.6 mg / mL, 0.8 mg / mL, and 1.0 mg / mL, respectively. These EP tubes were then cleaned in an ultrasonic cleaner for 15 minutes to ensure uniform distribution of the MnO2 nanoparticles in the suspension. Subsequently, the obtained MnO2 gradient suspensions were sequentially and uniformly added dropwise along a specified radius to the surface of an activated Blu-ray disc, forming corresponding sample droplets. After all the water in each sample droplet had completely evaporated, BD error readings were performed to obtain the data reading errors corresponding to the MnO2 gradient suspensions. The LDC of each signal peak was calculated, and grayscale analysis was conducted. The results of these analyses are presented as photographs of the MnO2 gradient suspension, super-depth-of-field micrographs of sample drop points, and LDC distribution maps, as shown in Figure 7(a). Furthermore, the LDC values ​​of the sample drop points were plotted against the grayscale values ​​of the original solution to reveal the dependence of LDC values ​​and grayscale values ​​on solution concentration, as shown in Figure 7(b).

[0105] Generally, grayscale analysis yields more accurate results, but it is time-consuming and costly. Conversely, LDC-based analysis is faster and less expensive. Referring to the fitted curve of grayscale value versus solution concentration shown in Figure 7(b), it can be seen that the fitted curve of LDC count versus solution concentration accurately reflects the actual solution concentration. In other words, this embodiment can effectively improve detection efficiency and reduce detection costs while ensuring detection accuracy.

[0106] Based on the standard curve shown in Figure 7(b) for recording the relationship between MnO2 solution concentration and LDC count, this embodiment presents a quantitative detection method for MnO2 solution based on a "direct spotting-drying" strategy, specifically including:

[0107] After the optical disc is activated, the MnO2 solution to be tested is dropped onto a specified radius on the surface of the optical disc to form a sample dropping point; it is easy to understand that the diameter of the MnO2 solution to be tested is the same as the diameter of the standard solution to be dropped when calibrating the standard curve;

[0108] After the water in the sample droplet has completely evaporated, the data in the optical disc is read, the data read errors are obtained, and the integral area of ​​the signal peak of the data read error is calculated to obtain the corresponding LDC count.

[0109] The concentration corresponding to the calculated LDC count is determined according to the pre-calibrated standard curve, and the concentration of the MnO2 solution to be tested is obtained.

[0110] It should also be noted that if the relevant standard curve has been calibrated in advance, there is no need to calibrate it again; the test can be performed directly based on the calibrated standard curve.

[0111] Example 3:

[0112] A method for quantitative detection of biological samples based on optical discs is used to achieve quantitative detection of GSH glutathione solution.

[0113] GSH is a non-protein thiol formed by the combination of glutamic acid, cysteine, and glycine. In the human body, it acts as an antioxidant to maintain cellular redox homeostasis. GSH mainly exists in the reduced state in the human body and can be oxidized by MnO2 to generate oxidized glutathione (GSSG), as shown in Figure 8(b).

[0114] To determine the concentration of the GSH solution, this embodiment first reacted a series of GSH solutions of varying concentrations with an equal concentration of excess MnO2 (2 mg / mL) solution individually. After the mixtures had completely reacted, they were centrifuged, causing the remaining MnO2 to deposit at the bottom of the centrifuge tube. Then, the supernatant was removed, thus removing the GSH and MnO2 produced in the reaction. 2+ Then, deionized water is added to resuspend MnO2, thus obtaining a set of pure MnO2 gradient suspensions, as shown in Figure 8(a).

[0115] The resulting gradient suspension of pure MnO2 was uniformly dropped onto the surface of an activated Blu-ray disc along a specified radius to form sample droplets. Data was then read from the Blu-ray disc to obtain the corresponding data reading errors, as shown in Figure 8(c). Since GSH can directly react with MnO2, the higher the GSH content, the higher the amount of MnO2 consumed, resulting in a corresponding decrease in the turbidity of the MnO2 suspension, consistent with the result shown in Figure 8(c). Furthermore, transmission electron microscopy (TEM) imaging of the MnO2 suspension was performed before and after the reaction. The TEM images showed a decrease in the density of MnO2 particles after the reaction, further demonstrating the property of GSH to dissolve MnO2.

[0116] Next, we used a "drip-dry strategy" (2D strategy) to perform BD error readings on the obtained MnO2 suspension and calculated the LDC count for each signal peak. The results showed a negative correlation between the LDC count of residual MnO2 and GSH concentration, as shown in Figure 8(d). Further analysis indicated a good linear relationship between LDC count and GSH concentration in the range of 2-16 mM, specifically calculated as: LDC count = (-1.951 × [GSH] + 5.850) × 10⁻⁶. 3 linear fit R 2 =0.991, as shown in (e) of Figure 8.

[0117] Calculations show that the detection limit (LOD) of GSH in this method is approximately 0.447 mM, which preliminarily meets the requirements for GSH detection in normal human cells (typically in the range of 1-10 mM).

[0118] Based on the above relationship between GSH solution concentration and LDC count, this embodiment presents a quantitative detection method for GSH solution based on a "direct spotting-drying" strategy, specifically including:

[0119] The GSH solution to be tested was mixed with an excess of MnO2 solution to allow the GSH and MnO2 to react fully. The mixed solution was then centrifuged. The concentration of the MnO2 solution was the same as that used in the calibration, which was 2 mg / mL.

[0120] After removing the supernatant from the centrifuged mixed solution, deionized water is added to the remaining solution to obtain a new MnO2 suspension. This MnO2 suspension is then dropped onto the surface of the activated Blu-ray disc at a specified radius to form sample dropping points. It is easy to understand that the diameter of the MnO2 suspension being dropped is the same as the diameter of the standard solution being dropped when calibrating the formula.

[0121] After the water in the sample droplet has completely evaporated, the data in the optical disc is read, the data read errors are obtained, and the integral area of ​​the signal peak of the data read error is calculated to obtain the corresponding LDC count.

[0122] Substituting the LDC count into the above relationship between GSH solution concentration and LDC count, the solubility of the GSH solution to be tested is obtained.

[0123] This embodiment provides a new approach to GSH detection, and it is expected that experimental conditions and parameters can be further optimized in the future to further reduce the detection limit.

[0124] It's easy to understand that when the selected MnO2 solution changes, the relevant fitting equation will also change; recalibration can then be performed according to the corresponding calibration method. Similarly, it should be noted that if the relevant equation has already been calibrated, no further calibration is needed; detection can be performed directly based on the calibrated equation.

[0125] Example 4:

[0126] A method for quantitative detection of biological samples based on optical discs, used to achieve quantitative detection of serum uric acid solution.

[0127] Serum uric acid is the final product of purine metabolism and is mainly excreted by the kidneys through urine. Under normal physiological conditions, the concentration of serum uric acid in men is usually in the range of 149-416 μmol / L (or 150-420 μmol / L), while in women it is 89-357 μmol / L (or 90-357 μmol / L).

[0128] Uric acid has reducing properties. When it comes into contact with manganese dioxide (MnO2), uric acid is oxidized, while MnO2 itself is reduced, producing allantoin and Mn. 2+ As shown in Figure 9, the detection principle of this embodiment is the same as that of Embodiment 3 above. To utilize the above-described "direct spotting-drying" strategy to detect serum uric acid content, this embodiment prepared a set of serum uric acid solutions with different concentration gradients and reacted them separately with an equal concentration of excess MnO2. After the reaction was complete, the mixed liquid was centrifuged, causing the remaining MnO2 to deposit at the bottom of the centrifuge tube. Subsequently, the supernatant was removed, thus removing allantoin and MnO2 generated during the reaction. 2+ Then, deionized water is added to resuspend the MnO2 suspension, thus obtaining a gradient suspension of pure MnO2. During this process, because serum uric acid reacts directly with MnO2, the higher the serum uric acid content, the higher the amount of MnO2 consumed, resulting in a corresponding decrease in the turbidity of the MnO2 suspension.

[0129] Next, the obtained MnO2 suspension was subjected to BD error readings, and the LDC count for each signal peak was calculated. These data were then converted into concentration curves for analysis and evaluation of serum uric acid levels. Similar to Example 3 above, this example calibrates the relationship between serum uric acid concentration and LDC count. Based on the calibrated relationship, this example describes a quantitative detection method for serum uric acid solution based on a "direct spotting-drying" strategy, specifically including:

[0130] The serum uric acid solution to be tested was mixed with an excess of MnO2 solution to allow the uric acid and MnO2 to react fully. The mixture was then centrifuged.

[0131] After removing the supernatant from the centrifuged mixed solution, deionized water is added to the remaining solution to obtain a new MnO2 suspension. This MnO2 suspension is then dropped onto the surface of the activated Blu-ray disc at a specified radius to form sample dropping points. It is easy to understand that the diameter of the MnO2 suspension being dropped is the same as the diameter of the standard solution being dropped when calibrating the formula.

[0132] After the water in the sample droplet has completely evaporated, the data in the optical disc is read, the data read errors are obtained, and the integral area of ​​the signal peak of the data read error is calculated to obtain the corresponding LDC count.

[0133] Substituting the LDC count into the above relationship between serum uric acid solution concentration and LDC count, the solubility of the GSH solution to be tested is obtained.

[0134] Example 5:

[0135] A method for quantitative detection of biological samples based on optical discs, used to achieve quantitative detection of organophosphorus pesticide solutions.

[0136] Current counting methods commonly employ traditional techniques such as capillary electrophoresis (CE), high-performance liquid chromatography (HPLC), and gas chromatography (GC) to achieve quantitative detection of organophosphorus pesticides (OPs). These methods have high detection sensitivity, but they rely on well-trained professionals and expensive laboratory equipment, making them unsuitable for on-site analysis.

[0137] Currently, a popular rapid on-site detection method combines colorimetric and enzyme inhibition methods. This method utilizes the catalytic hydrolysis of thioacetylcholine (ATCh) by acetylcholinesterase (AChE) to produce thiocholine (TCh), and the inhibition of AChE catalytic function by organophosphorus pesticides, thus affecting the subsequent colorimetric reaction. Based on this, this embodiment utilizes MnO2 as a common oxidant, reacting with thioacetylcholine (ATCh) to form water-soluble MnO2. 2+ As shown in Figure 10(a).

[0138] Since organophosphorus pesticides inhibit the catalytic function of acetylcholinesterase (AChE), their presence inhibits the dissolution of MnO2. Therefore, this embodiment can indirectly assess the concentration of organophosphorus pesticides by detecting the residual amount of MnO2. To utilize the "direct spotting-drying" strategy to detect the concentration of organophosphorus pesticide solutions, this embodiment first mixes a series of graded concentrations of organophosphorus pesticides with an equal concentration and excess of MnO2 suspension, thioacetylcholine solution, and acetylcholinesterase solution, allowing each mixture to react individually. After the mixtures have completely reacted, they are centrifuged, causing the remaining MnO2 to deposit at the bottom of the centrifuge tube. Next, the supernatant is removed, and deionized water is added to resuspend the MnO2 suspension, thus obtaining a series of pure MnO2 gradient suspensions. Since the presence of organophosphorus pesticides inhibits the dissolution of MnO2, the higher the content of organophosphorus pesticides, the higher the residual amount of MnO2, and the turbidity of the MnO2 suspension also increases, as shown in Figure 10(b).

[0139] The obtained gradient suspension of pure MnO2 was uniformly dropped onto the surface of an activated Blu-ray disc along a specified radius to form sample droplets. Data was then read from the Blu-ray disc to obtain corresponding data readout errors, and the LDC count for each signal peak was calculated. A positive correlation was found between the LDC count of residual MnO2 and the concentration of organophosphorus pesticides (OPs), as shown in Figure 10(c). Further analysis showed that the LDC count and OPs concentration were within the range of 1.289 × 10⁻⁶. -7 -1.289×10 -4 The LDC count exhibits an acceptable linear relationship within the range of M, specifically calculated as: LDC count = (0.608 × lg[OPs] + 4.386) × 10 3 linear fit R 2 =0.978, as shown in (d) of Figure 10. Furthermore, the detection limit for organophosphorus pesticides using this method is approximately 2.122 × 10⁻⁶. - 7 M meets actual testing needs.

[0140] It is easy to understand that when the selected MnO2 solution changes, the relevant fitting equation will also change, and recalibration can be performed according to the corresponding calibration method. Therefore, the dynamic range and detection limit can also be further optimized by adjusting the reaction time and MnO2 concentration.

[0141] Based on the above relationship between organophosphorus pesticide solution concentration and LDC count, this embodiment presents a quantitative detection method for organophosphorus pesticide solutions based on a "direct spotting-drying" strategy, specifically including:

[0142] The organophosphorus pesticide solution to be tested was mixed with an excess of MnO2 suspension, thioacetylcholine solution, and acetylcholinesterase solution. After the mixture was fully reacted, it was centrifuged.

[0143] After removing the supernatant from the centrifuged mixed solution, deionized water is added to the remaining solution to obtain a new MnO2 suspension. This MnO2 suspension is then dropped onto the surface of the activated Blu-ray disc at a specified radius to form sample dropping points. It is easy to understand that the diameter of the MnO2 suspension being dropped is the same as the diameter of the standard solution being dropped when calibrating the formula.

[0144] After the water in the sample droplet has completely evaporated, the data in the optical disc is read, the data read errors are obtained, and the integral area of ​​the signal peak of the data read error is calculated to obtain the corresponding LDC count.

[0145] Substituting the LDC count into the above relationship between the concentration of organophosphorus pesticide solution and the LDC count, the solubility of the organophosphorus pesticide solution to be tested is obtained.

[0146] It should also be noted that if the relevant relational expressions have already been calibrated, then no further calibration is needed; the detection can be performed directly based on the calibrated relational expressions.

[0147] Example 6:

[0148] A method for quantitative detection of biological samples based on optical discs, used to achieve quantitative detection of bacterial concentration.

[0149] Bacterial concentration detection plays an indispensable role in many key fields such as medicine, food, and environment, and is of great significance for the early detection of diseases, ensuring food safety, and assessing environmental hygiene. However, traditional bacterial detection methods are limited by their complexity and high cost, making it difficult to meet the needs of rapid on-site monitoring.

[0150] The detection principle of this embodiment is similar to that of embodiments 1 and 2 above. To achieve bacterial concentration detection based on the "direct spotting-drying" strategy, this embodiment first prepares a series of standard bacterial solutions with different concentrations by precise dilution with deionized water. Next, the Blu-ray disc undergoes a specific activation treatment, which changes the disc surface from hydrophobic to hydrophilic. Simultaneously, the ability of the disc surface to adsorb bacteria is enhanced after activation. Then, the standard bacterial solution to be tested is uniformly dropped onto the activated Blu-ray disc surface along a specified radius, forming sample droplets. As the water gradually evaporates, the bacteria in each sample droplet will deposit on the disc surface, forming clearly identifiable spotting points. Afterward, data is read from the Blu-ray disc to obtain the read data errors corresponding to each sample droplet. The LDC count of each signal peak in the error signal is further calculated. These counts reflect the density and distribution of bacterial deposition on the disc surface. Based on the calculated LDC counts, the concentration of each standard bacterial solution and the corresponding LDC count are fitted to obtain a standard curve for recording the correspondence between concentration and LDC count.

[0151] Based on the calibrated standard curve, this embodiment utilizes the "direct spotting-drying" strategy provided by the present invention for the quantitative detection method of bacterial solutions, specifically including:

[0152] After the optical disc is activated, a bacterial solution is dropped onto the surface of the optical disc at a specified radius to form a sample drop point. It is easy to understand that the diameter of the bacterial solution dropped is the same as the diameter of the standard solution dropped when calibrating the standard curve.

[0153] After the water in the sample droplet has completely evaporated, the data in the optical disc is read, the data read errors are obtained, and the integral area of ​​the signal peak of the data read error is calculated to obtain the corresponding LDC count.

[0154] The concentration of the bacterial solution is obtained by determining the concentration corresponding to the calculated LDC count according to the pre-calibrated standard curve.

[0155] The bacterial solution quantitative detection method provided in this embodiment is not only rapid but also highly accurate, and is applicable to various bacterial detection scenarios, providing an innovative method for rapid in vitro detection of bacteria.

[0156] Those skilled in the art will readily understand that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for quantitative detection of biological samples based on optical discs, used to detect the concentration of a test solution; wherein the test solution is a soluble target solution or an insoluble precipitate suspension, and neither the soluble target in the soluble target solution nor the insoluble precipitate in the insoluble precipitate suspension will damage the surface of the optical disc; The optical disc is filled with data; its characteristic is that... The quantitative detection method includes: after the optical disc is activated, the test solution is dropped onto a specified radius on the surface of the optical disc to form a sample drop point; the activation treatment causes the surface of the optical disc to change from hydrophobic to hydrophilic; after the water in the sample drop point has completely evaporated, the data in the optical disc is read, the data read errors are obtained, and the integral area of ​​the signal peak of the data read error is calculated to obtain the corresponding LDC count; the concentration corresponding to the calculated LDC count is determined according to a pre-calibrated standard curve to obtain the concentration of the test solution; wherein, when the test solution is a soluble target solution, the standard curve is obtained by fitting the LDC counts corresponding to soluble target solutions of different concentrations; when the test solution is an insoluble precipitate suspension, the standard curve is obtained by fitting the LDC counts corresponding to insoluble precipitate suspensions of different concentrations; the radius of the test solution added is the same as the radius of the liquid added when calibrating the standard curve.

2. The method for quantitative detection of biological samples based on optical discs as described in claim 1, characterized in that, The calibration method of the standard curve includes: preparing test solutions with different concentrations as standard solutions; after the optical disc is activated, uniformly adding the standard solutions along the specified radius on the surface of the optical disc to obtain sample drop points corresponding to each standard solution; after all the water in the sample drop points has completely evaporated, reading the data in the optical disc, obtaining the data reading errors corresponding to each sample drop point, and calculating the integral area of ​​the signal peak of each data reading error to obtain the LDC count corresponding to each standard solution; fitting the concentration of each standard solution and the corresponding LDC count to obtain a standard curve for recording the correspondence between concentration and LDC count.

3. A method for quantitative detection of biological samples based on optical discs, used to detect the concentration of a test solution, wherein the test solution is a soluble target solution; The optical disc is filled with data; its characteristic is that... The quantitative detection method includes: Step S1: Mixing the test solution with an excess of insoluble precipitate suspension, so that the soluble target in the test solution reacts fully with the insoluble precipitate in the insoluble precipitate suspension; the soluble target can react with the insoluble precipitate, and a soluble product is produced after the reaction; the insoluble precipitate will not damage the surface of the optical disc; Step S2: Centrifuging the mixed solution; Step S3: After removing the supernatant from the centrifuged mixed solution, adding deionized water to the remaining solution to obtain a new insoluble precipitate suspension; Step S4: After the optical disc is activated, the insoluble precipitate obtained in Step S3 is... The suspension is dropped onto a specified radius on the surface of the optical disc to form a sample droplet; the activation treatment causes the surface of the optical disc to change from hydrophobic to hydrophilic; Step S5: After the water in the sample droplet has completely evaporated, the data in the optical disc is read, the data read error is obtained, and the integral area of ​​the signal peak of the data read error is calculated to obtain the corresponding LDC count; Step S6: The LDC count obtained in step S5 is substituted into the pre-calibrated relationship between LDC count and the concentration of the test solution to obtain the concentration of the test solution; wherein, the radius of the insoluble precipitate suspension added is the same as the radius of the liquid added when calibrating the relationship between LDC count and the concentration of the test solution.

4. The method for quantitative detection of biological samples based on optical discs as described in claim 3, characterized in that, The calibration method for the relationship between LDC count and the concentration of the test solution includes: preparing test solutions with different concentrations as standard solutions; mixing each standard solution with an equal concentration and an excess of insoluble precipitate suspension to ensure sufficient reaction between the soluble target in each standard solution and the insoluble precipitate in the insoluble precipitate suspension; centrifuging each mixed solution separately; removing the supernatant from the centrifuged mixed solution and adding deionized water to the remaining solution to obtain a new insoluble precipitate suspension corresponding to each standard solution; after the optical disc is activated, uniformly adding the insoluble precipitate suspension corresponding to each standard solution along the specified radius on the surface of the optical disc to obtain sample dropping points corresponding to each standard solution; after all the water in the sample dropping points has completely evaporated, reading the data in the optical disc, obtaining the data reading errors corresponding to each sample dropping point, and calculating the integral area of ​​the signal peak of each data reading error to obtain the LDC count corresponding to each standard solution; fitting the concentration of each standard solution and the corresponding LDC count to obtain the relationship between the LDC count and the concentration of the test solution.

5. A method for quantitative detection of biological samples based on optical discs, used to detect the concentration of a test solution, wherein the test solution is a soluble target solution; The optical disc is filled with data; its characteristic is that... The quantitative detection method includes: Step T1: Mixing the test solution, the insoluble precipitate suspension, and the third reactant solution, so that the insoluble precipitate in the insoluble precipitate suspension reacts fully with the third reactant in the third reactant solution; the insoluble precipitate can react with the third reactant, and a soluble product is produced after the reaction; the insoluble precipitate suspension is in excess relative to the third reactant solution; the test solution inhibits the reaction between the third reactant and the insoluble precipitate, and the higher the concentration of the test solution, the stronger the inhibitory effect; the insoluble precipitate will not damage the surface of the optical disc; Step T2: Centrifuging the mixed solution; Step T3: After removing the supernatant from the centrifuged mixed solution, adding deionized water to the remaining solution to obtain a new insoluble precipitate solution. Step T4: After the optical disc is activated, the insoluble precipitate suspension obtained in step T3 is dropped onto a specified radius on the surface of the optical disc to form a sample droplet; the activation treatment changes the surface of the optical disc from hydrophobic to hydrophilic; Step T5: After the water in the sample droplet has completely evaporated, the data in the optical disc is read, the data read error is obtained, and the integral area of ​​the signal peak of the data read error is calculated to obtain the corresponding LDC count; Step T6: The LDC count obtained in step T5 is substituted into the pre-calibrated relationship between LDC count and the concentration of the test solution to obtain the concentration of the test solution; wherein, the radius of the insoluble precipitate suspension droplet is the same as the radius of the liquid added when calibrating the relationship between LDC count and the concentration of the test solution.

6. The method for quantitative detection of biological samples based on optical discs as described in claim 5, characterized in that, The calibration method for the relationship between LDC counting and the concentration of the test solution includes: preparing test solutions with different concentrations as standard solutions; for each standard solution, mixing it with an insoluble precipitate suspension and a third reactant solution to ensure sufficient reaction between the insoluble precipitate and the third reactant; ensuring that the concentrations of the insoluble precipitate suspensions mixed with each standard solution are equal, and that the concentrations of the third reactant solutions mixed with each standard solution are also equal; centrifuging each mixed solution separately; removing the supernatant from the centrifuged mixed solutions, adding deionized water to the remaining solutions to obtain new insoluble precipitates corresponding to each standard solution. Insoluble precipitate suspensions are prepared. After the optical disc is activated, the insoluble precipitate suspensions corresponding to each standard solution are uniformly added dropwise along the specified radius on the surface of the optical disc to obtain sample drop points corresponding to each standard solution. After all the water in the sample drop points has completely evaporated, the data in the optical disc is read to obtain the data reading errors corresponding to each sample drop point, and the integral area of ​​the signal peak of each data reading error is calculated to obtain the LDC count corresponding to each standard solution. The concentration of each standard solution and the corresponding LDC count are fitted to obtain the relationship between the LDC count and the concentration of the solution to be tested.

7. A method for quantitative detection of biological samples based on optical discs, used to detect the concentration of bacterial solutions; wherein the optical disc is filled with data; characterized in that, The quantitative detection method includes: after the optical disc is activated, the bacterial solution is dropped onto a specified radius on the surface of the optical disc to form a sample droplet; the activation treatment causes the surface of the optical disc to change from hydrophobic to hydrophilic; after the water in the sample droplet has completely evaporated, the data in the optical disc is read, the data read errors are obtained, and the integral area of ​​the signal peak of the data read error is calculated to obtain the corresponding LDC count; the concentration corresponding to the calculated LDC count is determined according to a pre-calibrated standard curve to obtain the concentration of the bacterial solution; wherein, the standard curve is obtained by fitting the LDC counts corresponding to different concentrations of bacterial solutions; the radius of the bacterial solution being dropped is the same as the radius of the liquid being dropped when calibrating the standard curve.

8. The method for quantitative detection of biological samples based on optical discs as described in claim 7, characterized in that, The calibration method for the standard curve includes: preparing bacterial solutions of different concentrations as standard solutions; after the optical disc is activated, uniformly adding the standard solutions along the specified radius on the surface of the optical disc to obtain sample drop points corresponding to each standard solution; after all the water in the sample drop points has completely evaporated, reading the data in the optical disc, obtaining the data reading errors corresponding to each sample drop point, and calculating the integral area of ​​the signal peak of each data reading error to obtain the LDC count corresponding to each standard solution; fitting the concentration of each standard solution and the corresponding LDC count to obtain a standard curve for recording the correspondence between concentration and LDC count.

9. The method for quantitative detection of biological samples based on optical discs as described in any one of claims 1 to 8, characterized in that, The activation treatment includes immersing the optical disc in NaOH solution for a preset time to generate reactive carboxyl groups on its surface, thereby changing it from a hydrophobic state to a hydrophilic state.

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