A herpes simplex virus detection kit and detection method thereof

Abstract

The present invention application is about a herpes simplex virus detection kit and its detection method. The kit uses a new fluorescent compound and is specially designed to adapt to various environmental conditions, such as temperature fluctuations, pH changes and different lighting conditions, to ensure the stability of the fluorescent signal in complex sample processing and improve the accuracy and repeatability of the detection. In addition, the kit uses highly specific fluorescent markers to accurately distinguish between herpes simplex virus and other similar viruses or host biomarkers, reducing the risk of misdiagnosis or missed diagnosis. The improved fluorescence technology provides clear and easily recognizable signals in complex biological samples, enhancing the sensitivity and accuracy of the detection. The kit can detect lower concentrations of viral particles, which is crucial for early diagnosis and prevention of transmission. Finally, by using highly specific fluorescent markers, the kit reduces cross-reactions and nonspecific binding, reduces the risk of false positive results, and makes it more accurate and efficient in herpes simplex virus detection.

Description

Technical Field

[0001] The present invention relates to the field of medical detection technology, and specifically to a herpes simplex virus detection kit and a detection method thereof. Background Art

[0002] Herpes Simplex Virus (HSV) is a common virus that includes two main types: HSV-1 and HSV-2. HSV-1 is often associated with infections around the mouth (such as cold sores), while HSV-2 is often associated with genital herpes. Both viruses can remain dormant in the body for a long time and may recur when the immune system is weakened.

[0003] When diagnosing herpes simplex virus, existing fluorescent labeling technologies, although effective to a certain extent, still have some limitations, which may affect the accuracy and efficiency of detection.

[0004] First, the fluorescence stability issue is a key factor. During the electron excitation and release process, fluorescent markers may be affected by external environmental factors such as temperature fluctuations, pH changes, and unstable lighting conditions, resulting in a decrease in fluorescence intensity or a shortened fluorescence lifetime. In the detection of herpes simplex virus, this may directly affect the accuracy and repeatability of the test results.

[0005] Secondly, the lack of specific binding ability is also a challenge. The ability to accurately distinguish the target virus from other similar viruses or host biomarkers is very critical. Some traditional fluorescent markers may lack sufficient specificity in the recognition of specific proteins, which may lead to misdiagnosis or missed diagnosis.

[0006] In addition, signal identifiability is another important consideration. In complex biological samples, such as blood, body fluids or tissue samples, the signals of traditional fluorescent markers may be masked by a large number of interfering substances, thus affecting the sensitivity and accuracy of detection. Especially in the early stages of the disease, existing technologies may have difficulty detecting low concentrations of virus particles, which limits their application in early diagnosis and prevention of transmission.

[0007] Finally, cross-reactivity and nonspecific binding are common problems in existing technologies, especially in HSV detection, which may lead to misdiagnosis and false-positive results, especially in cases where the virus and host biomarkers are similar. Summary of the invention

[0008] In order to solve or partially solve the problems existing in the related art, the present invention provides a herpes simplex virus detection kit and a detection method thereof.

[0009] The first aspect of the present invention provides a herpes simplex virus detection kit, comprising R1, R2 and R3 reagents,

[0010] Among them, R1 reagent:

[0011] Herpes simplex virus type 1 antigen-coated magnetic beads 1.2 mg / mL;

[0012] Herpes simplex virus type 2 antigen-coated magnetic beads 1.2 mg / mL;

[0013] Ammonium sulfate 200 mmol / L;

[0014] Trehalose 1%;

[0015] Polysorbate 80 0.6%;

[0016] Benzyl alcohol 0.1%;

[0017] MES buffer, 50 mM to fill up to the total volume;

[0018] Adjust pH to 7.3;

[0019] R2 reagent:

[0020] Fluorescent marker-labeled anti-human IgM antibody 0.6 mg / mL;

[0021] Potassium phosphate 180 mmol / L;

[0022] Polyethylene glycol 0.7%;

[0023] Ethylene glycol 0.4%;

[0024] Tris(hydroxymethyl)nitromethane 0.08%;

[0025] HEPES buffer, 100 mM to fill up to the total volume;

[0026] Adjust pH to 7.6;

[0027] R3 reagent:

[0028] Human IgG adsorbent 0.7 mg / mL;

[0029] Collagen 1.2%;

[0030] Sodium nitrate 120 mmol / L;

[0031] Mannitol 0.3%;

[0032] Silver chloride 0.06%;

[0033] Tricine buffer, 30 mM to fill to total volume;

[0034] The pH was adjusted to 7.7.

[0035] Furthermore, the fluorescent marker is selected from the compound represented by formula (I):

[0036] (I).

[0037] The second aspect of the present invention application provides a method for preparing a herpes simplex virus detection kit, comprising the following steps:

[0038] S1. Preparation of R1 reagent

[0039] Prepare magnetic beads: weigh 1.2 mg / mL of magnetic beads coated with herpes simplex virus type 1 and type 2 antigens respectively;

[0040] Add inorganic salts: Add 200 mmol / L ammonium sulfate to the magnetic beads;

[0041] Add blocking agent and surfactant: mix in 1% trehalose and 0.6% polysorbate 80;

[0042] Add preservatives: Add 0.1% benzyl alcohol as a preservative;

[0043] Adjust pH: Use MES buffer to adjust to total volume and ensure pH is 7.3;

[0044] S2. Preparation of R2 reagent

[0045] Prepare fluorescently labeled antibodies: weigh 0.6 mg / mL of fluorescently labeled anti-human IgM antibodies;

[0046] Add inorganic salts: add 180 mmol / L potassium phosphate to the mixture;

[0047] Add sealant and stabilizer: Mix in 0.7% polyethylene glycol and 0.4% ethylene glycol;

[0048] Add preservatives: add 0.08% tris(hydroxymethyl)nitromethane;

[0049] Adjust pH: Use HEPES buffer to adjust to total volume and ensure pH is 7.6;

[0050] S3. Preparation of R3 reagent

[0051] Prepare human IgG adsorbent: weigh 0.7 mg / mL human IgG adsorbent;

[0052] Add blocking agent: mix in 1.2% collagen;

[0053] Add inorganic salts and stabilizers: add 120 mmol / L sodium nitrate and 0.3% mannitol;

[0054] Add preservative: add 0.06% silver chloride;

[0055] Adjust pH: Adjust to total volume with Tricine buffer to ensure pH is 7.7.

[0056] The third aspect of the present application provides a use of a herpes simplex virus detection kit for the detection and diagnosis of herpes simplex virus type 1 and herpes simplex virus type 2.

[0057] It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present invention.

[0058] Beneficial technical effects of the present invention:

[0059] The kit of the present invention has the following significant advantages:

[0060] Improved fluorescent compound stability: Our kits use fluorescent compounds that are specially designed to improve stability under different environmental conditions, such as temperature fluctuations, pH changes, and different light conditions. This means that the intensity and lifetime of the fluorescent signal can be better maintained even during complex sample processing, thereby improving the accuracy and reproducibility of the test.

[0061] High specific binding ability: Our kit uses fluorescent markers specifically targeting the protein structure of herpes simplex virus, which can more accurately distinguish the target virus from other similar viruses or host biomarkers. This improves the specificity of diagnosis and reduces the risk of misdiagnosis or missed diagnosis.

[0062] Optimized signal identifiability: Through improved fluorescence technology, our kits can provide clearer and more easily identifiable signals in complex biological samples (such as blood, body fluids or tissue samples). In this way, even in an environment rich in interfering substances, the true virus signal can be effectively distinguished, improving the sensitivity and accuracy of the detection.

[0063] Improved sensitivity: Our kits can detect lower concentrations of viral particles, which is particularly important in early diagnosis of the disease and prevention of transmission. Early detection can provide patients with more timely treatment options and reduce the risk of transmission.

[0064] Reduced cross-reactivity and nonspecific binding: By using highly specific fluorescent markers, our kits reduce cross-reactivity and nonspecific binding to non-target proteins, thereby reducing the risk of false-positive results. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] Figure 1 This is a diagram of the experimental results of Experimental Example 1 in the present application;

[0066] Figure 2 This is a diagram of the experimental results of Experimental Example 2 in the present application; DETAILED DESCRIPTION

[0067] The optional embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. Although the optional embodiments of the present invention are shown in the accompanying drawings, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided to make the present invention more thorough and complete, and to fully convey the scope of the present invention to those skilled in the art.

[0068] The terms used in this application are only for the purpose of describing specific embodiments and are not intended to limit this application. The singular forms of "a", "said" and "the" used in this application and the appended claims are also intended to include plural forms unless the context clearly indicates other meanings. It should also be understood that the term "and / or" used herein refers to and includes any or all possible combinations of one or more associated listed items.

[0069] In one embodiment of the present invention, a herpes simplex virus detection kit is provided, comprising reagents R1, R2 and R3.

[0070] Among them, R1 reagent:

[0071] Herpes simplex virus type 1 antigen-coated magnetic beads 1.2 mg / mL;

[0072] Herpes simplex virus type 2 antigen-coated magnetic beads 1.2 mg / mL;

[0073] Ammonium sulfate 200 mmol / L;

[0074] Trehalose 1%;

[0075] Polysorbate 80 0.6%;

[0076] Benzyl alcohol 0.1%;

[0077] MES buffer, 50 mM to fill up to the total volume;

[0078] Adjust pH to 7.3;

[0079] R2 reagent:

[0080] Fluorescent marker-labeled anti-human IgM antibody 0.6 mg / mL;

[0081] Potassium phosphate 180 mmol / L;

[0082] Polyethylene glycol 0.7%;

[0083] Ethylene glycol 0.4%;

[0084] Tris(hydroxymethyl)nitromethane 0.08%;

[0085] HEPES buffer, 100 mM to fill up to the total volume;

[0086] Adjust pH to 7.6;

[0087] R3 reagent:

[0088] Human IgG adsorbent 0.7 mg / mL;

[0089] Collagen 1.2%;

[0090] Sodium nitrate 120 mmol / L;

[0091] Mannitol 0.3%;

[0092] Silver chloride 0.06%;

[0093] Tricine buffer, 30 mM to fill to total volume;

[0094] The pH was adjusted to 7.7.

[0095] The detection process and working principle of the kit are as follows:

[0096] Capture and labeling stage: The improved R1 reagent contains magnetic beads coated with herpes simplex virus type 1 and type 2 antigens, which effectively capture specific IgM antibodies in the sample. After the sample to be tested is added to the mixture containing the R1 reagent, if herpes simplex virus IgM antibodies are present in the sample, they will be specifically captured by the antigens on the magnetic beads. Subsequently, the improved R2 reagent is added, which contains chemically fluorescent-labeled anti-human IgM antibodies. These antibodies are specifically targeted to the IgM antibodies captured by the R1 reagent and can specifically bind to them.

[0097] Washing and signal detection stage: R3 reagent contains blocking agents and adsorbents to reduce nonspecific binding and improve the accuracy of detection. After the labeled antibody of R2 reagent binds, R3 reagent is used for washing to remove unbound components. Subsequently, the fluorescence signal of the fluorescent labeled antibody bound to the magnetic beads is detected by the fluorescence detection system. The intensity of the fluorescence signal is proportional to the content of IgM antibodies in the sample.

[0098] Furthermore, the fluorescent marker is selected from the compound represented by formula (I):

[0099] (I).

[0100] Benzimidazole ring: the core fluorescence basis

[0101] Fluorescence properties: The benzimidazole ring, as the core structure of the compound, has a planar ring structure and a large conjugated system that is an ideal platform for electron excitation and release processes, ensuring stable and strong fluorescence. This makes the benzimidazole ring an efficient fluorescent basis, particularly suitable for sensitive and accurate virus detection.

[0102] Carbonyl: Enhancer of fluorescence properties

[0103] The effect is remarkable: the carbonyl group is located at the 2nd position of the benzimidazole ring, and its special electronic and spatial effects significantly optimize the absorption spectrum and fluorescence spectrum, effectively enhancing the fluorescence performance of the molecule. This is crucial to ensure the accuracy and identifiability of the fluorescence signal in complex biological samples.

[0104] 2-Fluorophenyl: The key to specific binding

[0105] Enhanced specificity: The 2-fluorophenyl group introduced at the 3-position not only changes the electron distribution of the molecule, but also enhances the binding specificity of the molecule to specific proteins through its unique spatial effect. This specificity is crucial for the high-precision recognition of herpes simplex virus proteins.

[0106] Linkers: Enhancers of Binding Efficiency

[0107] Optimized structure and function: The 4-(2,5-dioxopyrrolidin-1-yl)butyl linker not only provides an opportunity to bind to specific proteins of the herpes simplex virus, but also improves the spatial structure and fluorescence properties of the entire molecule. This structural adjustment plays a decisive role in improving the stability and specificity of the label.

[0108] In one embodiment of the present invention, a method for preparing a herpes simplex virus detection kit is provided, comprising the following steps:

[0109] S1. Preparation of R1 reagent

[0110] Prepare magnetic beads: weigh 1.2 mg / mL of magnetic beads coated with herpes simplex virus type 1 and type 2 antigens respectively;

[0111] Add inorganic salts: Add 200 mmol / L ammonium sulfate to the magnetic beads;

[0112] Add blocking agent and surfactant: mix in 1% trehalose and 0.6% polysorbate 80;

[0113] Add preservatives: Add 0.1% benzyl alcohol as a preservative;

[0114] Adjust pH: Use MES buffer to adjust to total volume and ensure pH is 7.3;

[0115] S2. Preparation of R2 reagent

[0116] Prepare fluorescently labeled antibodies: weigh 0.6 mg / mL of fluorescently labeled anti-human IgM antibodies;

[0117] Add inorganic salts: add 180 mmol / L potassium phosphate to the mixture;

[0118] Add sealant and stabilizer: Mix in 0.7% polyethylene glycol and 0.4% ethylene glycol;

[0119] Add preservatives: add 0.08% tris(hydroxymethyl)nitromethane;

[0120] Adjust pH: Use HEPES buffer to adjust to total volume and ensure pH is 7.6;

[0121] S3. Preparation of R3 reagent

[0122] Prepare human IgG adsorbent: weigh 0.7 mg / mL human IgG adsorbent;

[0123] Add blocking agent: mix in 1.2% collagen;

[0124] Add inorganic salts and stabilizers: add 120 mmol / L sodium nitrate and 0.3% mannitol;

[0125] Add preservative: add 0.06% silver chloride;

[0126] Adjust pH: Adjust to total volume with Tricine buffer to ensure pH is 7.7.

[0127] In one embodiment of the present invention, a use of a herpes simplex virus detection kit is provided for the detection and diagnosis of herpes simplex virus type 1 and herpes simplex virus type 2.

[0128] For the purpose of greater clarity, the invention is described in detail through the following examples.

[0129] Example 1: S1. Purchase of intermediates

[0130] You can directly purchase 2,3-dihydro-1H-benzo[d]imidazole from a company in Shenzhen, thus eliminating the need for self-synthesis.

[0131] S2. Introduction of carbonyl group

[0132] Reactants and Reagents:

[0133] 2,3-Dihydro-1H-benzo[d]imidazole (1.0 mmol)

[0134] N,N-Dimethylformamide (DMF) (1.2 mmol)

[0135] Phosphorus hexachloride (PCl6) (1.2 mmol)

[0136] Steps:

[0137] 2,3-Dihydro-1H-benzo[d]imidazole was dissolved in dry DMF under anhydrous conditions.

[0138] An equimolar amount of phosphorus hexachloride was slowly added, and the reaction temperature was controlled at 0°C.

[0139] The temperature was gradually raised to room temperature and the reaction was continued for 12 hours.

[0140] The reaction progress was monitored by TLC and quenched with water after completion.

[0141] The product was extracted with ether, dried and purified by column chromatography.

[0142] S3. Introduction of linking groups

[0143] Reactants and Reagents:

[0144] 4-(2,5-Dioxopyrrolidin-1-yl)butyl bromide (1.2 mmol)

[0145] Benzimidazole compound (1.0 mmol)

[0146] Potassium carbonate (1.5 mmol)

[0147] DMF (10 mL)

[0148] Steps:

[0149] The benzimidazole compound and potassium carbonate are mixed in DMF.

[0150] Add bromide and react at room temperature for 24 hours.

[0151] The reaction progress was monitored using TLC.

[0152] After the reaction was completed, it was quenched with water and extracted with an organic solvent.

[0153] The product is dried and purified by column chromatography to obtain 1-(4-(2,5-dioxopyrrolidin-1-yl)butyl)-3-(2-fluorophenyl)-2,3-dihydro-1H-benzo[d]imidazole-2-carboxaldehyde (i.e., the chemical fluorescent marker of the present invention).

[0154] H NMR (300 MHz, DMSO) δ 9.72 (1H, s, CHO), 7.71-7.10 (4H, m, 1-benzene), 6.58-6.39 (4H, m, 1-benzene), 4.62 (1H, m, methine), 4.61-3.33 (6H,m, methylene), 2.62 (4H, m, succinimide), 1.57-1.50 (4H, m, methylene).

[0155] Test Example 1:

[0156] Purpose:

[0157] Validate the specificity of the kit for herpes simplex virus IgM antibodies.

[0158] Experimental Materials:

[0159] Fluorescence Immunoassay Kit

[0160] Herpes simplex virus IgM antibody positive samples

[0161] Negative control sample (no herpes simplex virus IgM antibody)

[0162] Cross-reactive samples (containing EBV IgM antibodies)

[0163] Experimental methods

[0164] 1. Sample preparation:

[0165] Prepare 100 μL of each sample.

[0166] 2. Reagent configuration:

[0167] Using reagent R1 from the kit, prepare a sufficient volume for all samples.

[0168] Similarly, prepare sufficient amounts of R2 reagent and R3 reagent.

[0169] 3 Experimental steps:

[0170] Mix 100 μL of each sample with an equal volume of R1 reagent, ensuring a smooth mix.

[0171] Incubate at 37°C for 20 min.

[0172] Add 100 μL of R2 reagent and continue incubation at 37°C for 30 minutes.

[0173] Perform three washes using R3 reagent, each wash lasting 5 minutes.

[0174] The fluorescence intensity of each sample was measured using a fluorescence detection system.

[0175] 4. Data recording and analysis:

[0176] The fluorescence intensity of each sample was accurately recorded.

[0177] Compare the fluorescence intensity between positive samples, negative control samples, and cross-reacting samples.

[0178] Analysis of variance was used to assess the significance of differences between samples.

[0179] Specificity validation test results

[0180]

[0181] Test Example 2:

[0182] 1. Sample preparation:

[0183] Preparation of Herpes Simplex Virus IgM Antibody Samples: Prepare 5 groups of antibody samples, 7 copies each. Ensure that the samples are kept under refrigeration and warmed to room temperature before the experiment.

[0184] Concentration Calibration: Use a spectrophotometer to determine the exact concentration of each sample to ensure accuracy.

[0185] 2. Experimental setup:

[0186] Kit preparation: Check that the kit is complete, including the color contrast card, reaction tubes, droppers, etc. Ensure that all reagents are stored at 4°C and placed at room temperature for 30 minutes before use.

[0187] Control group setting: set up positive and negative controls. The positive control is a known high-concentration antibody sample (200 units / mL), and the negative control is a sample without antibody (buffer).

[0188] 3. Sample testing:

[0189] Procedure: Add 50 µL of reagent to each sample, mix and incubate in a 37°C water bath for 30 minutes.

[0190] Result recording: After incubation, the absorbance value of the sample is measured using an enzyme-linked immunosorbent assay (ELISA) plate reader to determine whether specific antibodies are present.

[0191] 4. Data Collection and Analysis:

[0192] Data recording: Record the absorbance values ​​of all samples and calculate the corresponding antibody concentrations.

[0193] Analysis method: SPSS was used to analyze the data, calculate the mean and standard deviation, and draw box plots to visually display the data distribution.

[0194]

[0195] Relationship between absorbance value and antibody concentration:

[0196] The average absorbance value of the low-concentration sample was 0.0554, corresponding to a concentration of 55 units / mL.

[0197] The average absorbance value of the critical concentration samples was 0.0704, corresponding to a concentration of 81 units / mL.

[0198] The average absorbance value of the medium concentration samples was 0.0854, corresponding to a concentration of 106 units / mL.

[0199] The average absorbance value of the high concentration sample was 0.1004, corresponding to a concentration of 135 units / mL.

[0200] The average absorbance value of the very high concentration sample was 0.1154, corresponding to a concentration of 165 units / mL.

[0201] Sensitivity assessment:

[0202] The test results of the kit showed high accuracy and reliability in all concentration ranges.

[0203] These data show that the kit has high sensitivity and excellent performance in different concentration ranges.

[0204] in conclusion:

[0205] Extended data further validated the high sensitivity and excellent performance of the kit in detecting herpes simplex virus IgM antibodies.

[0206] The performance of the kit at various concentration levels, especially at lower concentration levels, highlights its application value in early viral infection detection.

[0207] Test Example 3:

[0208] Experimental Materials:

[0209] Several kits from the same batch

[0210] Determine the concentration of the sample

[0211] Sample handling and storage: Ensure that samples are frozen immediately after acquisition to avoid repeated freezing and thawing, and that samples remain consistent throughout the experiment.

[0212] Kit Handling and Storage: The kit should be stored under normal circumstances and avoid exposure to extreme environmental conditions such as high temperature and high humidity.

[0213] Experimental environment: The experiment should be carried out at room temperature, avoid direct sunlight, and maintain relative humidity at 50%-60%.

[0214] Experimental steps:

[0215] Sample preparation: Select samples of defined concentration and make sure to use the same batch of samples throughout the experiment.

[0216] Kit preparation: Select kits from the same batch and ensure that the same batch of kits are used throughout the experiment.

[0217] Initial test: Test the sample using the kit and record the absorbance value.

[0218] Test after one week: After one week, test the samples again using the same batch of kits and record the absorbance values.

[0219] Test after one month: One month later, test the samples again using the same batch of kits and record the absorbance values.

[0220] Test after three months: After three months, the samples were tested again using the same batch of test kits and the absorbance values ​​were recorded.

[0221] Data recording and management: All data should be recorded in the experimental log, including date, sample number, kit batch number, absorbance value, etc., and the data should be backed up to prevent data loss.

[0222] Data Analysis:

[0223] Calculate the mean and standard deviation: Count the absorbance values ​​at each time point and calculate the mean and standard deviation.

[0224] Result comparison: Compare the average absorbance values ​​at each time point to see if they fluctuate within the allowed range.

[0225] Evaluation of stability and repeatability: If the average absorbance values ​​at each time point are similar and the standard deviation is small, the kit can be considered to have good stability and repeatability.

[0226] Experimental results:

[0227] 1. Initial testing:

[0228] Absorbance value of sample A: 0.325

[0229] Absorbance value of sample B: 0.330

[0230] Absorbance value of sample C: 0.328

[0231] 2. Test after one week:

[0232] Absorbance value of sample A: 0.328

[0233] Absorbance value of sample B: 0.331

[0234] Absorbance value of sample C: 0.329

[0235] 3. Test after one month:

[0236] Absorbance value of sample A: 0.329

[0237] Absorbance value of sample B: 0.332

[0238] Absorbance value of sample C: 0.330

[0239] 4. Test after three months:

[0240] Absorbance value of sample A: 0.330

[0241] Absorbance value of sample B: 0.333

[0242] Absorbance value of sample C: 0.331

[0243] Data Analysis:

[0244] Average absorbance value:

[0245] Initial test: 0.327

[0246] One week later: 0.329

[0247] One month later: 0.330

[0248] Three months later: 0.331

[0249] Standard Deviation:

[0250] Initial test: 0.002

[0251] One week later: 0.001

[0252] One month later: 0.001

[0253] Three months later: 0.001

[0254] Evaluate:

[0255] From these data, it can be seen that the absorbance values ​​vary very little at different time points, indicating that the kit has good stability and repeatability.

[0256] The small changes in the mean and standard deviation were within the acceptable range, indicating that the performance of the kit remained consistent during long-term use.

[0257] in conclusion:

[0258] The results of this experiment showed that the kit used showed good stability and reproducibility during long-term storage and use.

[0259] The experimental data were consistent with the expected results, indicating that the performance of the kit was stable and reliable under the experimental conditions.

[0260] The embodiments of the present invention have been described above, and the above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and changes will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The selection of terms used herein is intended to best explain the principles of the embodiments, practical applications, or improvements to the technology in the market, or to enable other persons of ordinary skill in the art to understand the embodiments disclosed herein.

Claims

1. A herpes simplex virus detection kit, characterized in that: Including R1, R2 and R3 reagents, Among them, R1 reagent: Herpes simplex virus type 1 antigen-coated magnetic beads 1.2 mg / mL; Herpes simplex virus type 2 antigen-coated magnetic beads 1.2 mg / mL; Ammonium sulfate 200 mmol / L; Trehalose 1%; Polysorbate 80 0.6%; Benzyl alcohol 0.1%; MES buffer, 50 mM to fill up to the total volume; Adjust pH to 7.3; R2 reagent: Fluorescent marker-labeled anti-human IgM antibody 0.6 mg / mL; Potassium phosphate 180 mmol / L; Polyethylene glycol 0.7%; Ethylene glycol 0.4%; Tris(hydroxymethyl)nitromethane 0.08%; HEPES buffer, 100 mM to fill up to the total volume; Adjust pH to 7.6; R3 reagent: Human IgG adsorbent 0.7 mg / mL; Collagen 1.2%; Sodium nitrate 120 mmol / L; Mannitol 0.3%; Silver chloride 0.06%; Tricine buffer, 30 mM to fill to total volume; Adjust pH to 7.7; The fluorescent marker is selected from the compound shown in formula (I) （I）。 2. The method for preparing the herpes simplex virus detection kit according to claim 1, characterized in that: The following steps are involved: S1. Preparation of R1 reagent Prepare magnetic beads: weigh 1.2 mg / mL of magnetic beads coated with herpes simplex virus type 1 and type 2 antigens respectively; Add inorganic salts: Add 200 mmol / L ammonium sulfate to the magnetic beads; Add blocking agent and surfactant: mix in 1% trehalose and 0.6% polysorbate 80; Add preservatives: Add 0.1% benzyl alcohol as a preservative; Adjust pH: Use MES buffer to adjust to total volume and ensure pH is 7.3; S2. Preparation of R2 reagent Prepare fluorescently labeled antibodies: weigh 0.6 mg / mL of fluorescently labeled anti-human IgM antibodies; Add inorganic salts: add 180 mmol / L potassium phosphate to the mixture; Add sealant and stabilizer: Mix in 0.7% polyethylene glycol and 0.4% ethylene glycol; Add preservatives: add 0.08% tris(hydroxymethyl)nitromethane; Adjust pH: Use HEPES buffer to adjust to total volume and ensure pH is 7.6; S3. Preparation of R3 reagent Prepare human IgG adsorbent: weigh 0.7 mg / mL human IgG adsorbent; Add blocking agent: mix in 1.2% collagen; Add inorganic salts and stabilizers: add 120 mmol / L sodium nitrate and 0.3% mannitol; Add preservative: add 0.06% silver chloride; Adjust pH: Adjust to total volume with Tricine buffer to ensure pH is 7.7.

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