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Method for improving sensitivity and linearity of latex reagent

A sensitivity and latex technology, applied in the field of medical testing, can solve the problems of poor control of sensitivity and linearity, low measurement accuracy, waste of reagents, etc., to improve detection precision and accuracy, improve sensitivity and Linear relationship, high stability effect

Active Publication Date: 2019-05-17
浙江夸克生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] According to the above description, it can be seen that changes in body functions will correspond to changes in antigens in the body. In order to detect changes in various antigens, commonly used detection methods include fluorescent immunochromatography, colloidal gold, enzyme-linked immunosorbent assay, and latex-enhanced Although immunoassays, fluorescence immunochromatography, colloidal gold, and enzyme-linked immunosorbent assays can partially solve the sensitivity and linearity problems at the same time, they can only be qualitative or semi-quantitative, and the measurement accuracy is low; the commonly used latex-enhanced immunoassay ratio Nephelometric determination is accurate, but cannot take into account high sensitivity and broad line shape relationship
[0008] Latex immunoturbidimetry is divided into transmission turbidimetry and scattering turbidimetry. The former is used in biochemical analyzers, and the latter is used in specific protein analyzers. The kit needs to prepare different latex particles, and the sensitivity and linear relationship are not easy to control, so it is more troublesome to use. For example, to detect anti-streptolysin O, it is necessary to prepare goat anti-human anti-streptolysin O-conjugated latex particles; When detecting lipoprotein (a), it is necessary to prepare goat anti-human lipoprotein (a) antibody latex particles, which limits the detection of multiple antigens and easily causes waste of reagents
[0009] Therefore, based on the problems of strong detection pertinence, poor control of sensitivity and linear relationship in existing detection methods, a method that can be used to detect multiple antigens is provided, and the addition of this method can significantly improve latex detection. sensitivity and linearity, it is necessary

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  • Method for improving sensitivity and linearity of latex reagent
  • Method for improving sensitivity and linearity of latex reagent
  • Method for improving sensitivity and linearity of latex reagent

Examples

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Embodiment 1

[0059] Basic Example 1 A method for improving latex reagent sensitivity and linearity

[0060] Include the following steps:

[0061] (1) Preparation of solution A: the solution A includes a monoclonal antibody to the antigen to be tested connected to polystyrene microspheres;

[0062] The method for linking the monoclonal antibody against the antigen to be tested with the latex microspheres is: after washing the latex microspheres with 2-morpholineethanesulfonic acid buffer solution, the ethyl dimethyl Aminopropylcarbodiimide and the washed latex microspheres are mixed at room temperature for the first step reaction, and the reaction time is 1 hour. The latex microspheres after the first step reaction and the adipic acid dihydrazide solution According to the ratio of parts by weight of 80:30, they were mixed and reacted overnight at 5°C for the second step of reaction to obtain activated latex microspheres. Mix at a ratio of 5:100 and react overnight at 5°C for the third ste...

Embodiment 2

[0071] Basic Example 2 A latex turbidimetric detection kit for preparing an antibody to be detected, the kit includes reagent R1 and reagent R2. The volume ratio of the reagent R1 and the reagent R2 is 3:1.

[0072] The preparation method of described reagent R2 comprises:

[0073] A. Add bovine serum albumin of 1-10g / L to the prepared C solution, and react;

[0074] B, add the glucose of 1-8g / L in the solution that above-mentioned step (1) reacts to obtain, react;

[0075] C, the solution that has reacted in the ammonium chloride buffer solution washing step (2), washes 2-3 times, then adds the sucrose of 0.5-5g / L, adjusts the latex microsphere concentration to be 0.15-0.20% with buffer solution, namely Reagent R2 is obtained.

[0076] The reaction described in the above step A is to react at 4-8°C for 1-2 hours; the reaction condition described in the above step B is to react overnight at 4-8°C.

[0077] The weight-number ratio of the bovine serum albumin in the above st...

Embodiment 3

[0089] Example 3 A kind of latex turbidimetric detection kit for preparing and detecting glycocholic acid

[0090] Detection reagents include R1 and R2,

[0091] The R1 reagent is: trismethylolaminomethane 100mmol / L, polyethylene glycol 6000 4g / L, sodium chloride 15g / L.

[0092] The R2 reagent is: polycaprolactone latex microspheres sensitized with 0.18% anti-glycocholic acid antibody in 0.1M Tris buffer.

[0093] The volume ratio of the reagent R1 and the reagent R2 is 4:1.

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Abstract

The invention provides a method for improving sensitivity and linearity of a latex reagent, and belongs to the field of medical test. The method comprises the following steps: step (1), preparing a solution A: the solution A includes a monoclonal antibody connected to a latex microsphere for an antigen to be tested; step (2), preparation a solution B: the solution B includes phenylalanine, lysineand vitamin C; and step (3), mixing the solution A obtained in step (1) and the solution B obtained in step (2) to obtain a solution C, wherein the solution C is used for detecting the antigen to be tested. The invention further provides a preparation method of a detection kit, wherein a certain amount of the solution B is added to a reagent R2, and the addition of the solution B not only improvesthe sensitivity of the detection, but also can obviously improve the linear relationship of the detection; and the addition of the solution B is unexpectedly found to improve the stability of the reagent R2, and the stability of the reagent R2 is still relatively high under the condition that a preservative does not need to be added.

Description

technical field [0001] The invention belongs to the field of medical detection, in particular to a method for improving the sensitivity and linearity of latex reagents. Background technique [0002] Apolipoprotein E (Apolipoprotein E, APOE) is a polymorphic protein that participates in the transformation and metabolism of lipoproteins. Its genes can regulate many biological functions and are related to the pathogenesis of many ophthalmic diseases. The study of APOE is one of the hotspots in medical research at present. To explore the inner relationship between APOE and its gene polymorphism has important clinical application value for the prevention, diagnosis and treatment of eye diseases. Normal human plasma APOE concentration is 0.03-0.05g / L. ApoE concentration is positively correlated with plasma triglyceride content. [0003] Antistreptolysin O (ASO) is one of the metabolites of group A streptococci, which can dissolve human hemoglobin and has strong antigenicity. To...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/531G01N33/543
Inventor 陈青松林耀文
Owner 浙江夸克生物科技有限公司
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