阅读说明：本技术 胶乳增强免疫比浊法试剂盒 (Latex enhanced immunoturbidimetry kit ) 是由 陈开华 于 2019-09-09 设计创作，主要内容包括：本发明提供了一种胶乳增强免疫比浊法试剂盒,其包括第一试剂、第二试剂和校准品；其中,第一试剂包括缓冲液、稳定剂、促凝剂、表面活性剂和防腐剂；第二试剂包括包被有抗体的胶乳颗粒、缓冲液、稳定剂、表面活性剂和防腐剂；校准品包括抗原、缓冲液、稳定剂、表面活性剂和防腐剂；本发明提供的试剂盒与目前临床使用试剂盒相比,特异性、灵敏度和准确性一致,且本发明的试剂盒适用于一般检测机构都具有的生化分析仪上使用,检测时间短,操作方便,检测成本低,无放射性污染,更适合临床推广使用。(The invention provides a latex enhanced immunoturbidimetry kit, which comprises a first reagent, a second reagent and a calibrator; wherein the first reagent comprises a buffer, a stabilizer, a coagulant, a surfactant and a preservative; the second reagent comprises latex particles coated with the antibody, a buffer solution, a stabilizing agent, a surfactant and a preservative; the calibrator comprises antigen, buffer solution, stabilizer, surfactant and preservative; compared with the existing clinical kit, the kit provided by the invention has the advantages of consistent specificity, sensitivity and accuracy, short detection time, convenience in operation, low detection cost, no radioactive pollution and suitability for clinical popularization and use, and is suitable for biochemical analyzers of common detection mechanisms.)

1. A latex enhanced immunoturbidimetry kit is characterized in that: the kit comprises a first reagent, a second reagent and a calibrator;

the first reagent comprises a buffer, a stabilizer, a coagulant, a surfactant and a preservative;

the second reagent comprises latex particles coated with antibodies, a buffer, a stabilizer, a surfactant and a preservative;

the calibrator comprises an antigen, a buffer solution, a stabilizer, a surfactant and a preservative.

2. The latex-enhanced immunoturbidimetry kit of claim 1, wherein: the latex particles are polystyrene latex particles;

the surface modification of the polystyrene latex particles is selected from more than one of sulfate group, sulfonic group, carboxyl group, amino group, hydroxyl group or chloromethyl group;

the diameter of the latex particles is 60-200nm, and the concentration is 0.08-0.5%.

3. The latex-enhanced immunoturbidimetry kit of claim 1, wherein: the buffer solution is selected from more than one of Tris buffer solution, Tris-HCl buffer solution, phosphate buffer solution or glycine buffer solution.

4. The latex-enhanced immunoturbidimetry kit of claim 1, wherein: the surfactant is selected from more than one of Tween40, TritonX-100 or Span 80.

5. The latex-enhanced immunoturbidimetry kit of claim 1, wherein: the stabilizer is selected from more than one of bovine serum albumin with the concentration range of 0.1-5%, sodium chloride with the concentration range of 0.1-5%, EDTA with the concentration range of 2-50mM, Tween-20 with the concentration range of 0.1-1% or glycerol with the concentration range of 1-10%.

6. The latex-enhanced immunoturbidimetry kit of claim 1, wherein: the preservative is selected from more than one of sodium azide, potassium sorbate, phenol or sodium nitrite.

7. The latex-enhanced immunoturbidimetry kit of claim 1, wherein: the coagulant is selected from more than one of PEG-4000, PEG-6000, PEG-8000, PEG-10000, dextran sodium sulfate, dextran T-40 or polybrene.

8. A method of preparing the latex-enhanced immunoturbidimetric kit of any of claims 1-7, wherein: which comprises the following steps:

(1) respectively preparing a first reagent and a calibrator;

(2) reacting the latex particles with the antibody to coat the antibody; the coated latex is dispersed in a buffer solution after being washed and sealed to prepare a second reagent;

(3) and uniformly mixing the first reagent, the second reagent and the calibrator to obtain the latex enhanced immunoturbidimetry kit.

Technical Field

The invention belongs to the technical field of in-vitro diagnosis of medical immunity, and particularly relates to a latex enhanced immunoturbidimetry kit.

Background

A latex particle-enhanced turbidimetric immunoassay (PETIA) is a relatively stable and accurate method for detecting the homogeneous immunoturbidimetric assay of the humoral protein in recent years. The PETIA method is mainly divided into two methods, one is a scattering turbidimetric assay, the other is a projection turbidimetric assay, the basic principles of the two methods are very similar, monoclonal antibodies are crosslinked on the surfaces of polymer latex microspheres, and after the microspheres crosslinked with the antibodies are combined with antigens, the monoclonal antibodies can be rapidly aggregated together in a short time, so that the light scattering performance or the light transmittance performance of a reaction solution is changed. Moreover, the change of the light-scattering property or the light-transmitting property (i.e., the absorbance) of the reaction solution has strong correlation with the concentration of the antigen to be detected, and the concentration of the antigen to be detected can be reflected in a certain range. The PETIA detection method is to perform antigen and antibody reactions and result measurement in a homogeneous reaction system. After the antigen and the antibody react, the absorbance value of the reaction solution is directly measured, the complex operation steps of repeatedly incubating and washing a plate by an ELISA method are omitted, the result can be obtained in a few minutes, and time and labor are saved. In addition, the simplification of the operation steps of the nano immunoturbidimetry correspondingly avoids the interference of a plurality of human operation factors and external factors such as reagents, environment and the like, has better stability and repeatability, and can reflect the content of the substance to be detected more truly. Although the sensitivity of the immunoturbidimetry is poorer than that of the ELISA method, the lower limit value of the immunoturbidimetry is enough for detecting a plurality of marker proteins in the blood plasma of healthy people, and the clinical detection requirements can be completely met.

Disclosure of Invention

Aiming at the defects in the prior art, the invention aims to provide a latex enhanced immunoturbidimetry kit.

In order to achieve the above purpose, the solution of the invention is as follows:

a latex-enhanced immunoturbidimetry kit comprising a first reagent, a second reagent, and a calibrator;

the first reagent comprises a buffer, a stabilizer, a coagulant, a surfactant and a preservative;

the second reagent comprises latex particles coated with antibodies, a buffer, a stabilizer, a surfactant and a preservative;

the calibrator comprises an antigen, a buffer solution, a stabilizer, a surfactant and a preservative.

Preferably, the latex particles are polystyrene latex particles.

Preferably, the surface modification of the polystyrene latex particles is one or more selected from the group consisting of a sulfate group, a sulfonate group, a carboxyl group, an amino group, a hydroxyl group and a chloromethyl group.

Preferably, the latex particles have a diameter of 60 to 200nm and a concentration of 0.08 to 0.5%.

Preferably, the buffer is selected from more than one of Tris buffer, Tris-HCl buffer, phosphate buffer or glycine buffer.

Preferably, the surfactant is selected from more than one of Tween40, TritonX-100 or Span 80.

Preferably, the stabilizer is selected from more than one of bovine serum albumin with the concentration range of 0.1-5%, sodium chloride with the concentration range of 0.1-5%, EDTA with the concentration range of 2-50mM, Tween-20 with the concentration range of 0.1-1% or glycerol with the concentration range of 1-10%.

Preferably, the preservative is selected from one or more of sodium azide, potassium sorbate, phenol or sodium nitrite.

Preferably, the coagulant is selected from more than one of PEG-4000, PEG-6000, PEG-8000, PEG-10000, dextran sodium sulfate, dextran T-40 or polybrene.

The preparation method of the latex-enhanced immunoturbidimetric kit comprises the following steps:

(1) respectively preparing a first reagent and a calibrator;

(2) reacting the latex particles with the antibody to coat the antibody; and (3) dispersing the coated latex in a buffer solution after the steps of washing and blocking to prepare a second reagent.

(3) And uniformly mixing the first reagent, the second reagent and the calibrator to obtain the latex enhanced immunoturbidimetry kit.

Due to the adoption of the scheme, the invention has the beneficial effects that:

compared with the existing clinical kit, the kit provided by the invention has the advantages of consistent specificity, sensitivity and accuracy, short detection time, convenience in operation, low detection cost, no radioactive pollution and suitability for clinical popularization and use, and is suitable for biochemical analyzers of common detection mechanisms.

Detailed Description

The present invention will be further described with reference to the following examples.