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Homocysteine detection kit and homocysteine detection method

A technology of homocysteine ​​and detection kits, which is applied in biological testing, measuring devices, material inspection products, etc., and can solve problems such as complex operation, low sensitivity of cyclic enzyme method, and high sample requirements of enzyme-linked immunosorbent method

Pending Publication Date: 2022-07-29
深圳天辰医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the methods used to detect Hcy mainly include liquid high-performance liquid phase method, cyclic enzyme method, enzyme-linked immunoassay and chemiluminescence immunoassay. low sensitivity; enzyme-linked immunoassay has high requirements on samples and low sensitivity

Method used

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  • Homocysteine detection kit and homocysteine detection method
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  • Homocysteine detection kit and homocysteine detection method

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0046] The present invention also provides a preparation method of the above-mentioned homocysteine ​​detection kit, comprising the following steps:

[0047] (1) Preparation of calibrators

[0048] The calibrator dilution was prepared in 50 mM Tris buffer pH 7.4 containing: 50 mM Tris, 100 mM NaCl, 1.2% BSA, 1 g / L Proclin 300. According to the set concentration of Hcy calibrator, the corresponding pure Hcy was added to the above buffer to prepare 5 calibrators with a series of concentrations; the concentrations of the 5 calibrators were 0 μmol / L, 1 μmol / L, 5 μmol / L, 10μmol / L, 20μmol / L and 50μmol / L.

[0049] (2) Preparation of the first reagent

[0050] a. Take 10 mg of tosyl magnetic beads and resuspend them in 0.1M, pH 9.5 boric acid buffer, place on a magnetic stand for 1 minute, and remove the supernatant;

[0051] b. Add 900 μL of 0.1M boric acid buffer, pH 9.5, and mix by vortexing;

[0052] c. Add the corresponding amount of SAH-coated antibody to the final reaction ...

Embodiment 1

[0073] Example 1 Detection of Sensitivity

[0074]Referring to the experimental protocol recommended in the CLSI EP17-A document, 5 low-value samples with concentrations similar to the detection limit were tested, and each sample was tested 5 times. The number of detection results (blank limit is 0.5μmol / L) should be less than or equal to 3, it can be considered that the settings of blank limit and detection limit provided by the manufacturer are basically reasonable, and the sensitivity is 1μmol / L. It shows that the homocysteine ​​detection kit provided by the present invention has higher sensitivity.

[0075] Table 1 Sensitivity test result table

[0076] serial number S1 S2 S3 S4 S5 1 0.71 0.82 0.80 0.88 0.91 2 0.69 0.71 0.82 0.89 0.95 3 0.61 0.74 0.76 0.76 0.90 4 0.66 0.83 0.85 0.84 0.97 5 0.68 0.75 0.73 0.82 0.86

Embodiment 2

[0077] Embodiment 2 Linear detection

[0078] Linear analysis was performed on calibrators with concentrations of 0 μmol / L, 1 μmol / L, 5 μmol / L, 10 μmol / L, 20 μmol / L and 50 μmol / L, and the results are shown in Table 2 and figure 1 , the linear correlation coefficient r=0.9981 was calculated, and the linear range of the kit was 1-50 μmol / L. This shows that the homocysteine ​​detection kit provided by the present invention has a wide linear range.

[0079] Table 2 Linear analysis results of calibrators

[0080] Concentration (μmol / L) Luminescence value (RLU value) 0 1302952 1 1255639 5 1112153 10 985784 20 762603 50 52254

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Abstract

The invention discloses a homocysteine detection kit and a homocysteine detection method.The homocysteine detection kit comprises a first reagent, a second reagent, a third reagent and a fourth reagent, the first reagent is a magnetic particle working solution coated with an SAH antibody, and the second reagent is a magnetic particle working solution coated with an SAH antibody; the kit comprises a first reagent, a second reagent, a third reagent and a fourth reagent, the first reagent is alkaline phosphatase labeled SAH derivative enzyme labeled working solution, the second reagent is alkaline phosphatase labeled SAH derivative enzyme labeled working solution, the third reagent is a reducing agent buffer solution, the reducing agent is at least one of DTT, TCEP and beta-mercaptoethanol, and the fourth reagent is a buffer solution of SAHH and Ado. The homocysteine detection kit provided by the invention has the advantages of high sensitivity, accurate quantification, no radioactive risk, simplicity in operation, short detection time and difficulty in cross reaction.

Description

technical field [0001] The invention relates to the technical field of in vitro detection, in particular to a homocysteine ​​detection kit and a homocysteine ​​detection method. Background technique [0002] Homocysteine ​​(Hcy) is one of the three sulfur-containing amino acids in the body, an intermediate product of methionine metabolism, and itself is not involved in protein synthesis. The methionine molecule contains an S methyl group; after interacting with ATP to generate S-adenosylmethionine, it can provide methyl groups for about 50 known substances with important physiological activities in the body through various transmethylation reactions. After S-adenosylmethionine transfers the methyl group to another substance under the action of methyltransferase, S-adenosyl-homocysteine ​​is generated, and the latter is further removed from adenosine to generate homocysteine. About 70% of homocysteine ​​in plasma is bound to albumin and is a bound type. The rest of the free...

Claims

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Application Information

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IPC IPC(8): G01N21/76G01N33/543G01N33/68
CPCG01N21/76G01N33/543G01N33/6893G01N2800/042G01N2800/321G01N2800/32G01N2800/368G01N2800/28G01N2800/085G01N2800/347
Inventor 何裕勇黄金浪唐灿
Owner 深圳天辰医疗科技有限公司
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