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A latex-enhanced immunoturbidimetric assay kit for detecting creatine kinase isoenzyme ck-mb

A technology of latex enhancement and immune turbidimetry, which is applied in the field of immunoanalysis medicine, can solve the problems of detection interference, achieve the effect of reducing detection cost, wide detection range, and satisfying clinical use

Active Publication Date: 2022-04-15
SHENZHEN AMTECH BIOENGINEERING LTD INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the inventors found in their research that when the latex-enhanced immunoturbidimetric method was applied to the detection of CK-MB in blood samples, the rheumatoid factor (rheumatoid factor, RF) present in the samples would interfere with the detection
In addition, the non-specific binding of proteins and other components in the sample to the antibody leads to background signals and interferes with the detection

Method used

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  • A latex-enhanced immunoturbidimetric assay kit for detecting creatine kinase isoenzyme ck-mb
  • A latex-enhanced immunoturbidimetric assay kit for detecting creatine kinase isoenzyme ck-mb
  • A latex-enhanced immunoturbidimetric assay kit for detecting creatine kinase isoenzyme ck-mb

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] The kit of this embodiment includes R1 reagent and R2 reagent.

[0063] The formula of R1 reagent is as follows:

[0064]

[0065] The preparation method of R1 reagent is as follows:

[0066] Weigh 1g PEG10000, 1.5g rabbit anti-mouse IgM antibody, 0.2g Tween-40, 2g BSA and 0.2g sodium azide, add to 100mL phosphate buffer (pH 5), stir well to promote dissolution, mix evenly, and obtain R1 reagent.

[0067] The formula of R2 reagent is as follows:

[0068]

[0069] R2 reagent preparation method is as follows:

[0070] Weigh 1g of polystyrene microspheres with an average particle size of 200nm and carboxyl groups on the surface, dilute it to a concentration of 1% (w / v) with a pH 5 phosphate buffer, add an appropriate amount of 1-(3-dimethylamino Propyl)-3-ethylcarbodiimide hydrochloride (EDC) solution, activated at 4°C for 1 hour, centrifuged at 20,000 rpm for 15 minutes, removed the supernatant, and added pH 5 phosphate buffer to the precipitate , ultrasonicall...

Embodiment 2

[0077] The kit of this embodiment includes R1 reagent and R2 reagent, the formula of which is as follows, and the preparation method refers to Example 1. The kit can also be equipped with CK-MB concentrations of 0ng / mL, 5ng / mL, 10ng / mL, 20ng / mL, 40ng / mL, 100ng / mL, 300ng / mL calibrator, and CK-MB concentrations of 5- 50ng / mL and 50-200ng / mL quality controls.

[0078] The formula of R1 reagent is as follows:

[0079]

[0080] The formula of R2 reagent is as follows:

[0081]

[0082]

Embodiment 3

[0084]The kit of this embodiment includes reagent R1 and reagent R2, the formula of which is as follows, and the preparation method refers to Example 1. The kit can also be equipped with CK-MB concentrations of 0ng / mL, 5ng / mL, 10ng / mL, 20ng / mL, 40ng / mL, 100ng / mL, 300ng / mL calibrator, and CK-MB concentrations of 5- 50ng / mL and 50-200ng / mL quality controls.

[0085] The formula of R1 reagent is as follows:

[0086]

[0087] The formula of R2 reagent is as follows:

[0088]

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Abstract

The invention provides a latex-enhanced immune turbidimetric assay kit for detecting creatine kinase isoenzyme CK-MB, comprising R1 reagent and R2 reagent, wherein the R1 reagent comprises a buffer, a coagulant, a blocking agent, and a surfactant , protective agent, preservative, R2 reagent contains buffer, CK‑MB antibody-coated nanospheres, protective agent, and preservative. In Reagent 1, rabbit anti-mouse IgM antibody or SeaBlock fish plasma blocking agent was used as blocking agent, and in Reagent 2, CK‑MB antibody-coated nanospheres were used as three components of glycine + ethanolamine + calf serum as blocking agent , which can effectively reduce detection interference. The kit of the present application has the advantages of strong anti-interference, high sensitivity, wide detection range, good repeatability, high specificity, and good stability. It can realize automatic detection on a biochemical analyzer, replace chemiluminescent products, and reduce detection costs. Meet clinical use.

Description

technical field [0001] The invention relates to the technical field of immunoanalysis medicine, in particular to a latex-enhanced immune turbidimetry kit for detecting creatine kinase isoenzyme CK-MB. Background technique [0002] Creatine kinase (CK) is divided into cytoplasmic type and mitochondrial type. The cytoplasmic type is divided into three subtypes: CK-MM, CK-MB, and CK-BB according to the difference between M subunit and B subunit. Two mt subunits make up CK-mt. CK-MB is a hybrid creatine kinase isoenzyme, which is a form of creatine kinase, mainly exists in cardiomyocytes, and is one of the markers of myocardial injury. [0003] CK-MB has been recommended as the gold standard for the diagnosis of acute myocardial infarction (AMI) due to its good specificity and sensitivity. However, since there is also a small amount of CK-MB in skeletal muscle, the CK-MB in skeletal muscle in children will be higher, so the specificity is not good, and there is a risk of misdi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/573G01N33/557G01N33/543
CPCG01N33/573G01N33/557G01N33/54346G01N33/54393G01N2333/9123G01N2800/324
Inventor 陈小茹吴向东严小莉
Owner SHENZHEN AMTECH BIOENGINEERING LTD INC