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Myoglobin assay kit

A technology for detecting kits and myoglobin, which is applied in the field of medical immunology, can solve the problems of increasing the amount of antibodies and production costs, weak anti-interference ability, and false positive test results, so as to improve accuracy and reliability, and resist Strong interference ability and the effect of reducing the amount of antibodies

Active Publication Date: 2014-05-14
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] And antibody is coated on the latex, generally can adopt physical adsorption method and chemical coupling method, the sensitization latex particle stability that adopts physical adsorption method to obtain is poor, and antibody comes off easily from latex particle; , may be interfered by rheumatoid factor (RF) and heterophile antibodies, resulting in false positive or falsely elevated test results
At present, most of the chemical coupling methods used are random coupling methods, because the randomly coupled antibodies will be randomly coupled to different parts of latex organisms, which will lead to the loss of binding force of the antibodies, increasing the amount of antibodies and production costs; if the above method The obtained sensitized latex particles are used in detection kits, which will cause defects of poor specificity, low accuracy, weak anti-interference ability or high production cost

Method used

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Experimental program
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Effect test

Embodiment 1

[0028] The present invention will be described in further detail below through examples, but the present invention is not limited to the following examples. Example 1 Preparation of MB detection kit

[0029] The main raw materials of the reagent kit of the present invention are as follows:

[0030] 1. MB antibody: polyclonal antibody (commercially available).

[0031] 2. Latex: The present invention only exemplarily uses polystyrene latex particles (commercially available) with a diameter of 150-250 nm with carboxyl groups for experiments.

[0032] The main reagents of this embodiment are prepared as follows:

[0033] Reagent R1: Glycine buffer containing 1.5% PEG6000 (polyethylene glycol 6000), 95mmol / L NaCl, this reagent is a colorless and transparent solution.

[0034] Reagent R2: sensitize polystyrene latex particles with a particle size of 200 nm with anti-human MB antibody. The reagent is a milky white solution. Specific steps are as follows:

[0035] 1. Take 1ml (100mg / ml) latex...

Embodiment 2

[0045] Example 2 Determination of MB

[0046] Testing tool: Hitachi 7060 automatic analyzer.

[0047] Analytical method: two-point endpoint method; main wavelength: 570nm, sub-wavelength: 800nm: sample volume: 6ul (microliter); R1: 200ul; R2: 50ul; reaction direction: rising; measurement temperature: 37℃; sample mixed with R1 After homogenization, read the absorbance A at the 10th second 1 ;Add R2 at 3~5min, read absorbance A after 5min 2 . The absorbance of the reaction is calculated as A 2 With A 1 The difference.

[0048] Calculation method: Multi-point calibration, draw a dose / response curve based on the absorbance and the value of the reference serum, and the sample content can be calculated on the dose / response curve based on its absorbance value.

Embodiment 3

[0049] Example 3 Performance Evaluation of MB Detection Kit

[0050] 1. Evaluation of analytical sensitivity

[0051] Use 5% bovine serum albumin solution as a blank sample, and the blank sample should not contain the analyte. Perform 20 consecutive tests on the biochemical analyzer, and calculate the mean and standard deviation SD of the 20 results. The blank mean plus twice the standard deviation (+2SD) was used as the detection limit of the reporting method. It can be seen from Table 1 that the sensitivity is 1.59ug / L.

[0052] Table 1

[0053]

[0054] 2. High-value linear evaluation

[0055] Press 1 part of low-value serum (20ug / L) and 1 part of high-value serum (800ug / L) according to 9:1, 4:1, 7:3, 3:2, 1:1, 2:3, 3:7 , 1:4, 1:9 (the sample sequence number is 2-10, and the sample 1 is the low-value serum before mixing), and prepare 10 samples with different concentrations. Repeat the measurement twice for each sample. Table 2 shows that the highest detection range of the detec...

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Abstract

The invention discloses a myoglobin assay kit. The myoglobin assay kit is characterized in that the kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 is of an appropriate buffer solution; and the reagent R2 is formed by placing myoglobin antibody-sensitized polystyrene latex particles in the appropriate buffer solution. The myoglobin assay kit has the advantages of simple and fast detection, high sensitivity, good accuracy, strong anti-interference ability and low production cost.

Description

Technical field [0001] The present invention belongs to the field of medical immunology, and relates to an immunological detection reagent. Further, the present invention relates to a myoglobin detection kit. Background technique [0002] Myoglobin (MB) is a specific oxygen-binding protein for skeletal muscle and myocardium, with a molecular weight of 17.8kD. Due to its small molecular weight, when myocardium or skeletal muscle is damaged, MB will be released from the muscle tissue into the circulating blood and can It is filtered by the glomerulus and appears in the urine. Therefore, the measurement of MB in blood and urine can be used for the diagnosis of certain myopathies and heart diseases, such as acute myocardial infarction (AMI), acute muscle injury, acute and chronic renal failure, severe congestive heart failure, prolonged shock, muscular dystrophy, Diseases such as muscle atrophy and polymyositis. [0003] AMI is a serious life-threatening disease. The main biochemical...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
Inventor 邹炳德邹继华
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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