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Myoglobin determination kit of compound antibody

A technology for myoglobin and human myoglobin, which is applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of short reaction time, the detection sensitivity and linear range need to be further improved, and the precision is good, and the accuracy is achieved. High performance, improved detection sensitivity and good precision

Inactive Publication Date: 2015-04-22
JIANGSU HAOSHEN MEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The PETIA detection method has short reaction time, good precision, wide detection range, and is not affected by jaundice, hemolysis and lipemia specimens. It is easy to automate and is currently a very widely used detection method. However, the detection sensitivity and linear range of this method still needs to be further improved

Method used

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  • Myoglobin determination kit of compound antibody
  • Myoglobin determination kit of compound antibody
  • Myoglobin determination kit of compound antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Preparation of myoglobin assay kit

[0023] 1. Preparation of Reagent R1

[0024] Add final concentrations of 0.5% polyethylene glycol 6000, 0.1% Tween 20, 0.1% bovine serum albumin, and 0.1% sodium azide to 20 mmol / L phosphate buffer (pH 7.4), and fully Mix well, and filter with a 0.22 μm filter membrane to prepare reagent R1.

[0025] 2. Preparation of Reagent R2

[0026] Mouse anti-human myoglobin monoclonal antibody and rabbit anti-human myoglobin polyclonal antibody are both entrusted to professional companies to produce according to conventional methods. The antibody only reacts with human myoglobin and has no immune cross-reaction with other antigens. Meet the needs of this test.

[0027] Add EDAC:Sulfo-NHS:nanolatex particles (250nm or 60nm) to 20mmol (pH6.1) NES buffer solution, the mass ratio is 1:0.6:1, mix well, activate on a rotary shaker at room temperature for 1 hour, and centrifuge at 17,000rpm After 30min, wash with 20mmol (pH6.1) NES buffer thre...

Embodiment 2

[0030] Performance Test of Myoglobin Assay Kit

[0031] Experimental analysis method: two-point endpoint method. Take 150 ul of reagent R1, add 25 ul of sample, add 50 ul of reagent R2 after 5 minutes at 37°C, start to read the point, react for 5 minutes, read the point again, and obtain the absorbance difference (△A).

[0032] Testing instrument: Hitachi 7180 automatic analyzer.

[0033] Detection wavelength: 500nm.

[0034] 1. Sensitivity test

[0035] Detect water 20 times, record the absorbance difference (△A 水 ), calculate the mean value (X 水 ) and standard deviation (SD 水 ); detect samples with a concentration of 100μg / L 20 times, and record the absorbance difference (△A 样 ), calculate the mean value (X 样 ) and standard deviation (SD 样 ), and the test results are shown in Table 1. Calculate the sensitivity using the following formula.

[0036] Sensitivity = 100 × (X 水 +3×SD 水 ) / X 样

[0037] Table 1

[0038]

[0039] The sensitivity of the reagent...

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Abstract

The invention provides a myoglobin determination kit of a compound antibody. The myoglobin determination kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 is a reaction liquid for promoting antigen specific binding of the antibody; the reagent R2 is a buffer solution of nano latex micro-particles combined with an anti-human-myoglobin antibody; and the reagent R2 is characterized by being prepared by virtue of the following steps: coupling polystyrene nano latex micro-particles of which the diameters are 50-300nm with a myoglobin monoclonal antibody or a polyclonal antibody to obtain conjugates with different particle sizes and different antibodies; and mixing the conjugates with different particle sizes and different antibodies according to different proportions to prepare the reagent R2. The reagent R2 can be used for detecting myoglobin in human serum, and can be used for improving the detection sensitivity and linear range of the kit provided by the invention. The detection sensitivity of the kit is 0.2mu g / L, and the linear range of the kit is 0-1200mu g / L.

Description

technical field [0001] The invention belongs to the field of medical testing, and in particular relates to a nano latex microparticle type myoglobin assay kit. Background technique [0002] Myoglobin (Mbglobin, Mb) is a heme protein composed of a polypeptide chain of 153 amino acids and a heme prosthetic group, with a relative molecular mass of 17500, mainly distributed in striated muscle (cardiac and skeletal muscle) cells . Due to its small molecular weight, Mb is the best early marker for entering the blood when cardiomyocytes are damaged. Mb is filtered through the glomerulus and excreted in the urine, so the determination of the concentration of Mb in serum and urine can be used for the diagnosis of some myopathy and heart disease. Such as acute myocardial infarction, acute and chronic renal failure, severe congestive heart failure, prolonged shock, neuromuscular disease and myopathy caused by various reasons. [0003] Acute myocardial infarction (AMI) is a common cl...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/54346G01N33/6887G01N2333/805G01N2800/324G01N2800/50
Inventor 董同义李玲董海江张文静
Owner JIANGSU HAOSHEN MEDICAL TECH
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