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Latex enhanced immunoturbidimetry kit for detecting creatine kinase isoenzyme CK-MB

A latex enhancement, immune turbidimetric technology, applied in the field of immunoassay medicine, can solve problems such as detection interference, achieve the effects of reducing detection cost, strong anti-interference, and reducing false positive results

Active Publication Date: 2020-08-21
SHENZHEN AMTECH BIOENGINEERING LTD INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Latex enhanced immunoturbidimetry kit for detecting creatine kinase isoenzyme CK-MB
  • Latex enhanced immunoturbidimetry kit for detecting creatine kinase isoenzyme CK-MB
  • Latex enhanced immunoturbidimetry kit for detecting creatine kinase isoenzyme CK-MB

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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 it 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 1 g of polystyrene microspheres with an average particle size of 200 nm and carboxyl groups on the surface, dilute it to a concentration of 1% (w / v) with 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 , ultrasonic...

Embodiment 2

[0077] 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.

[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 immunoturbidimetry kit for detecting creatine kinase isoenzyme CK-MB. The latex enhanced immunoturbidimetry kit comprises a reagent R1 and a reagent R2, whereinthe reagent R1 comprises a buffer solution, a coagulant, a blocking agent, a surfactant, a protective agent and a preservative, and the reagent R2 comprises a buffer solution, CK-MB antibody coated nano microspheres, a protective agent and a preservative. A rabbit anti-mouse IgM antibody or a SeaBlock fish plasma blocking agent is adopted as the blocking agent in the reagent 1, and glycine, ethanolamine and calf serum are adopted as a sealing agent in the CK-MB antibody coated nano microspheres of the reagent 2, so that the detection interference can be effectively reduced. The kit provided by the invention has the advantages of strong anti-interference performance, high sensitivity, wide detection range, good repeatability, high specificity, good stability and the like, can realize automatic detection on a biochemical analyzer, replace a chemiluminescence product and reduce the detection cost, and meets 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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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
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