A method and a kit for detecting salivary uric acid

Abstract

The application discloses a saliva uric acid detection method and a detection kit, belongs to the field of in-vitro diagnosis, and aims to solve the problem that there is no mature commercial uric acid non-invasive detection product at present. Technical scheme points are as follows: an excess of an oxidizing agent is added to react with uric acid, when uric acid is too high, the oxidizing agent is completely consumed, at this time, the mixed solution of saliva and the reaction reagent does not react with the detection test strip or the detection reagent, the detection test strip does not develop color, or after the detection reagent is added to the mixed solution of saliva and the reaction reagent, the mixture does not develop color, which indicates high uric acid. The uric acid detection method provided by the application can well distinguish normal uric acid and high uric acid, and the method is simple, convenient, mature commercial, and can become a simple non-invasive self-detection method for a patient with hyperuricemia to pay attention to the uric acid level change of the patient at any time and for a long time.

Description

Technical Field

[0001] This invention relates to the field of in vitro diagnostics, and more specifically, to a method and reagent kit for detecting sialic acid. Background Technology

[0002] Hyperuricemia is a chronic metabolic disease caused by purine metabolism disorders. In its early stages, it generally presents with no obvious symptoms. If patients do not pay enough attention to it and fail to detect, control, or treat it effectively in time, it will develop into gout, leading to clinical manifestations such as acute arthritis and tophi. It may also be accompanied by obesity, type 2 diabetes, dyslipidemia, hypertension, arteriosclerosis, and coronary heart disease. Therefore, it is especially important for high-risk individuals to monitor their uric acid levels and control them within the ideal range.

[0003] Currently, the routine clinical method for detecting uric acid, whether at a hospital or at home, involves blood testing. Blood testing provides the most direct reflection of blood uric acid levels, but hospital testing is relatively cumbersome, and home testing also requires blood collection, which places a certain psychological burden on patients. Both methods result in poor patient compliance and are not conducive to patients monitoring changes in their uric acid levels regularly and over the long term.

[0004] Existing studies have shown a positive correlation between sialic acid and blood uric acid, making sialic acid testing a novel way to reflect one's own uric acid levels. Saliva collection is convenient and non-invasive, offering significant advantages over blood collection. Therefore, convenient methods for detecting sialic acid are increasingly becoming a research hotspot; however, no mature, commercially available sialic acid testing products have yet emerged.

[0005] Therefore, a solution needs to be proposed to address this problem. Summary of the Invention

[0006] To overcome the inconvenience of existing blood uric acid testing methods, this invention provides a low-cost, convenient, and rapid non-invasive salivary uric acid detection method and kit.

[0007] This invention achieves the above objective through the following technical solution: a method for detecting sialic acid, characterized by comprising the following steps: S1: Take 1 part of saliva into a container, add 0.4 parts of oxidation reaction reagent, shake, and let stand for 1-2 minutes; S2: Insert the redox test strip into the container to contact the liquid in S1 after it has been left to stand, and then remove it. Observe the color change of the redox test strip. Alternatively, add the redox detection reagent into the container and mix it thoroughly with the liquid in S1 after it has been left to stand. Observe the color change of the mixture.

[0008] The present invention is further configured such that the oxidation reaction reagent is a solution containing an oxidizing reagent.

[0009] The present invention is further configured such that the oxidizing agent includes, but is not limited to, one or more compounds having permanganate, manganate, dichromate, chromate, periodate, iodate, hypochlorite, chlorate, hypobromate, bromate, pyrophosphate, and hydrogen peroxide.

[0010] The present invention is further configured such that the solution containing the oxidizing agent also contains a non-oxidizing-reducing compound, including but not limited to one or more of water, sodium chloride, potassium chloride, sodium acetate, potassium acetate, acetic acid, phosphoric acid, sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, potassium phosphate, potassium hydrogen phosphate, potassium dihydrogen phosphate, boric acid, and tris(hydroxymethyl)aminomethane.

[0011] The present invention is further configured such that the concentration of the oxidizing agent in the solution is 0.01%~1%.

[0012] The present invention is further configured such that the concentration of the non-redox compound in the solution is 0~1M.

[0013] The present invention is further configured such that: the redox test strip is a test strip for detecting oxidizing reagents, and the redox detection reagent is a reagent for detecting oxidizing reagents.

[0014] A kit for detecting salivary uric acid includes a saliva collector, a collection tube, a graduated reaction tube, the aforementioned oxidation reaction reagent, a container containing the oxidation reaction reagent, the aforementioned redox test strip or redox detection reagent and its container, and a plastic sampling pipette.

[0015] Compared with the prior art, the beneficial effects of the present invention are: Firstly, this invention utilizes the reducing properties of uric acid. It involves adding an excess of the reaction equivalent of an oxidizing agent (compared to normal uric acid levels) to react with the uric acid. When uric acid is too high, the oxidizing agent is completely consumed. At this point, the mixture of saliva and the reaction agent shows no color reaction with the redox test strip or redox detection reagent, indicating high uric acid. When uric acid is normal, the oxidizing agent is not completely consumed. The excess oxidizing agent in the mixture of saliva and the reaction agent reacts with the redox test strip or redox detection reagent, indicating normal uric acid. The entire operation is very convenient, and the required reagents are easy to prepare and inexpensive. Secondly, saliva also contains many other reducing substances. The reaction reagents provided by the preferred embodiment of the present invention can reduce or eliminate the influence of other reducing substances besides uric acid, so as to avoid false positive results. Attached Figure Description

[0016] Figure 1 This is a schematic diagram of the results of Experiment 1; Figure 2 This is a schematic diagram of the results of Experiment 2; Figure 3 This is a schematic diagram of the results of Experiment 3; Figure 4 This is a schematic diagram of the results of Experiment 4. Figure 1 ; Figure 5 This is a schematic diagram of the results of Experiment 4. Figure 2 . Detailed Implementation

[0017] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. In this description, it should be understood that the terms "upper," "lower," "front," "rear," "left," "right," "top," "bottom," "inner," and "outer," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing the present 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. Therefore, they should not be construed as limitations on the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Example

[0018] A method for detecting sialic acid includes the following steps: S1: Take 1 part of saliva into a container, add 0.4 parts of oxidation reaction reagent, shake, and let stand for 1-2 minutes; S2: Insert the redox test strip into the container to contact the liquid in S1 after it has been left to stand, and then remove it. Observe the color change of the redox test strip. Alternatively, add the redox detection reagent into the container and mix it thoroughly with the liquid in S1 after it has been left to stand. Observe the color change of the mixture.

[0019] If the user needs to change the upper limit of saliva uric acid detection, this can be achieved by increasing or decreasing the amount of oxidation reaction reagent. For example, in the current embodiment, 100ul is mixed with 40ul of 1mg / ml sodium periodate aqueous solution, which corresponds to an upper limit of blood uric acid value of about 480. If, for some monitoring needs, the upper limit of monitoring needs to be adjusted to about 400, then the amount of saliva used can be reduced while keeping the amount of oxidation reaction reagent unchanged.

[0020] The oxidizing agent includes, but is not limited to, compounds containing permanganate, manganate, dichromate, chromate, periodate, iodate, hypochlorite, chlorate, hypobromate, bromate, pyrophosphate, and one or more of hydrogen peroxide, preferably compounds containing periodate and iodate.

[0021] The solution containing the oxidizing agent also contains non-oxidizing and reducing compounds, including but not limited to one or more of water, sodium chloride, potassium chloride, sodium acetate, potassium acetate, acetic acid, phosphoric acid, sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, potassium phosphate, potassium hydrogen phosphate, potassium dihydrogen phosphate, boric acid, and tris(hydroxymethyl)aminomethane.

[0022] The concentration of the oxidizing agent in the solution is 0.01% to 1%, preferably 0.05% to 0.4%.

[0023] The solution contains non-redox compounds at a concentration of 0-1 M, preferably 0-300 mM.

[0024] The redox test strip is a test strip for detecting oxidizing reagents, preferably a starch-potassium iodide test strip; the redox detection reagent is a reagent for detecting oxidizing reagents, preferably a starch-potassium iodide solution.

[0025] A kit for detecting saliva uric acid includes a saliva collector and collection tube, a graduated reaction tube, an oxidation reaction reagent and a container containing the oxidation reaction reagent, an oxidation-reduction test strip or oxidation-reduction detection reagent and its container, and a plastic sampling pipette.

[0026] Experiment 1: Simulated Detection of Salivary Uric Acid 1 Uric acid was dissolved in commercially available PBS buffer to prepare uric acid solutions with concentrations of 100 μg / ml, 70 μg / ml, and 50 μg / ml. A sodium iodate solution was also prepared using PBS to achieve a concentration of 1 mg / ml. 0.1 ml of each of the 100 μg / ml, 70 μg / ml, and 50 μg / ml solutions were placed in clean 2 ml centrifuge tubes. 20 μl of the prepared sodium iodate solution was added to each tube, and the mixture was manually shaken thoroughly. After standing for 1 minute, a commercially available starch-potassium iodide test strip was immersed in the mixture. The strip was immediately removed after contact with the solution, and the color change was observed. The results are as follows: Figure 1 As shown, the higher the uric acid concentration, the lighter the color of the starch-potassium iodide test strip, which is in line with expectations. Furthermore, the color difference trend between uric acid solutions of different concentrations is consistent with the concentration difference.

[0027] Example 2: Simulated detection of saliva uric acid 2 Uric acid was dissolved in deionized water to prepare uric acid solutions with concentrations of 100 μg / ml and 50 μg / ml, respectively. A sodium periodate solution was also prepared using deionized water to achieve a concentration of 1 mg / ml. 0.1 ml of each of the 100 μg / ml and 50 μg / ml solutions were placed in clean 2 ml centrifuge tubes. 40 μl of the prepared sodium periodate solution was added to each tube, and the mixture was manually shaken thoroughly. After standing for 1 minute, a commercially available starch-potassium iodide test strip was immersed in the mixture. The strip was immediately removed after contact with the solution, and the color change was observed. The results are as follows: Figure 2 As shown: the higher the uric acid concentration, the lighter the color, which is as expected.

[0028] Example 3: Real Saliva Test One male with normal uric acid, one female with normal uric acid, and two males with high uric acid (uric acid level above 500) were selected. After oral cleaning, approximately 0.5 ml of naturally flowing saliva was collected, and 0.1 ml of each was placed into four clean 2 ml centrifuge tubes. A sodium periodate solution with a concentration of 1 mg / ml was prepared using deionized water. Four 40 μl portions of the sodium periodate solution were added to the four 2 ml centrifuge tubes containing saliva. The tubes were manually shaken to mix thoroughly. After standing for 2 minutes, a commercially available starch-potassium iodide test strip was inserted into the test strip and immediately removed. The color change of the starch-potassium iodide test strip was observed. The results are as follows: Figure 3 As shown: In the diagram, m represents a male with normal uric acid levels, f represents a female with normal uric acid levels, and M1 and M2 represent two males with high uric acid levels. The starch-potassium iodide test strip results show that the samples from both male and female individuals with normal uric acid levels show distinct color changes, with the female samples showing a darker color because women generally have lower uric acid levels than men. However, the samples from the two males with high uric acid levels show no color change at all, which is consistent with expectations. This demonstrates that the uric acid detection method provided by this invention effectively distinguishes between normal and high uric acid levels.

[0029] Example 4: Tracking one's own uric acid levels One female with normal uric acid, one male with normal uric acid, and one male with high uric acid (uric acid level consistently above 500) were selected. The female with normal uric acid ate normally within 18 hours before the test, the male with normal uric acid consumed a large amount of seafood within 18 hours before the test, and the male with high uric acid had been controlling his diet for 3 months and ate normally within 18 hours before the test. After cleaning their mouths, approximately 0.5 ml of naturally flowing saliva was collected, and 0.1 ml of each was placed in three clean 2 ml centrifuge tubes. A sodium periodate solution with a concentration of 1 mg / ml was prepared using deionized water. Three 40 μl portions of the sodium periodate solution were added to the three 2 ml centrifuge tubes containing saliva, and the mixture was manually shaken to mix thoroughly. After standing for 2 minutes, a commercially available starch-potassium iodide test strip was inserted into the test strip and immediately removed. The color change of the starch-potassium iodide test strip was observed. The results are as follows. Figure 4As shown; one week later, saliva samples from the same three individuals were collected again for testing. The women and men with normal uric acid levels had normal diets in the 18 hours prior to the test, while the man with high uric acid had consumed meat broth in the 18 hours prior to the test. The results were as follows. Figure 5 As shown.

[0030] Figure 4 and Figure 5 In the diagram, L represents women with normal uric acid levels, H represents men with normal uric acid levels, and C represents men with high uric acid levels. The results show that... Figure 4 The L signal indicates that the uric acid level is normal, but the signal is weak, suggesting that the uric acid level may have fluctuated to near the upper limit of the normal range during the test. The H signal is absent, indicating that the uric acid level rose after consuming a large amount of seafood. The C signal is extremely weak, indicating that dietary control may be effective, but the uric acid level is still near the borderline, and it is necessary to continue to control the diet and monitor the uric acid level. Figure 5 The L and H signals in the test indicate that the uric acid level is normal. The strong H signal suggests that people with normal uric acid have a faster metabolic rate, and their uric acid level returns to normal quickly after their diet returns to normal. C showed no signal at all, indicating that his uric acid level was above the critical value. Due to his slow metabolic rate, even with some dietary fluctuations, the uric acid level recovered more slowly. The salivary uric acid test results obtained by the method of this invention suggest that man C with high uric acid still needs to pay attention to controlling his diet and changing his lifestyle. His blood uric acid level on the day of the test was 609, far higher than 500, and the blood uric acid test results also suggest that man C with high uric acid needs to control his diet and change his lifestyle. The results of this embodiment accurately reflect the individual's own uric acid level.

[0031] The above embodiments illustrate that the salivary uric acid detection method provided by the present invention can conveniently, quickly, and accurately reflect whether one's own uric acid level is normal. It can become a simple and non-invasive self-testing method, especially for patients with hyperuricemia, to monitor changes in their own uric acid level at any time and over a long period of time.

[0032] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

[0033] Furthermore, it should be understood that although this specification describes embodiments, not every embodiment contains only one independent technical solution. This narrative style is merely for clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can also be appropriately combined to form other embodiments that can be understood by those skilled in the art.

Claims

1. A method for detecting sialic acid, characterized in that, Includes the following steps: S1: Take 1 part of saliva into a container, add 0.4 parts of oxidation reaction reagent, shake, and let stand for 1-2 minutes; S2: Insert the redox test strip into the container to contact the liquid in S1 after it has been left to stand, and then remove it. Observe the color change of the redox test strip. Alternatively, add the redox detection reagent into the container and mix it thoroughly with the liquid in S1 after it has been left to stand. Observe the color change of the mixture.

2. The method for detecting sialic acid according to claim 1, characterized in that: The oxidation reaction reagent is a solution containing an oxidizing reagent.

3. The method for detecting sialic acid according to claim 2, characterized in that: The oxidizing agent includes, but is not limited to, compounds containing permanganate, manganate, dichromate, chromate, periodate, iodate, hypochlorite, chlorate, hypobromate, bromate, pyrophosphate, and one or more of hydrogen peroxide.

4. The method for detecting sialic acid according to claim 3, characterized in that: The solution containing the oxidizing agent also contains non-oxidizing and reducing compounds, including but not limited to one or more of water, sodium chloride, potassium chloride, sodium acetate, potassium acetate, acetic acid, phosphoric acid, sodium phosphate, sodium hydrogen phosphate, sodium dihydrogen phosphate, potassium phosphate, potassium hydrogen phosphate, potassium dihydrogen phosphate, boric acid, and tris(hydroxymethyl)aminomethane.

5. The method for detecting sialic acid according to claim 2, characterized in that: The concentration of the oxidizing agent in the solution is 0.01% to 1%.

6. The method for detecting sialic acid according to claim 5, characterized in that: The solution contains non-redox compounds at a concentration of 0-1 M.

7. The method for detecting sialic acid according to claim 1, characterized in that: The redox test strip is a test strip for detecting oxidizing reagents, and the redox detection reagent is a reagent for detecting oxidizing reagents.

8. A kit for detecting sialic acid, characterized in that: It includes a saliva collector, a collection tube, a graduated reaction tube, an oxidation reaction reagent as described in any one of claims 1 to 7, a container containing the oxidation reaction reagent, an oxidation-reduction test strip or oxidation-reduction detection reagent as described in claim 7 and its container, and a plastic sampling pipette.

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