Detection kit for measuring content of myohemoglobin in serum

A technology for detecting kits and myoglobin, which is applied in the field of medical immunological in vitro diagnosis, can solve the problems of expensive chemiluminescence method and enzyme-linked fluorescence analysis method, long detection cycle of enzyme-linked immunoassay method, and unsuitability for emergency rapid detection, etc. Achieve the effects of high accuracy of measurement results, easy operation and high sensitivity

Inactive Publication Date: 2013-11-13
上海睿康生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods have their limitations. For example, chemiluminescence and enzyme-linked fluorescence analysis methods are expensive, require special instruments and professionals to oper

Method used

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  • Detection kit for measuring content of myohemoglobin in serum
  • Detection kit for measuring content of myohemoglobin in serum
  • Detection kit for measuring content of myohemoglobin in serum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: The composition of assay kit is as follows:

[0032] 1. Reagent R1 is:

[0033] PH7.4 phosphate buffer 45mmol / l

[0034] Polyethylene glycol 600080mmol / l

[0035] Disodium EDTA 17mmol / l

[0036] NaN30.9%

[0037] Components can be added sequentially at room temperature, added simultaneously, or individually packaged and prepared just before assay.

[0038] 2. Reagent R2 is:

[0039] Myoglobin polyclonal antibody and polystyrene latex particles are mixed at a mass ratio of 30:100, and the buffer is 0.02M phosphate buffer (pH7.4). After the two are mixed evenly, they are adsorbed at 37°C for 8 hours, and then dialyzed Remove unlinked antibodies, add blocking solution (0.1% skimmed milk powder, 0.2mm glycine), and block for 2 hours. Centrifuge to remove the supernatant, dilute with latex (PH7.4 phosphate buffer 50mmol / l, disodium edetate 16mmol / l, 0.1% skimmed milk powder, 0.9%NaN3) to 0.25%.

[0040] 3. Calibrator preparation:

[0041] Recombinant ...

Embodiment 2

[0044] Embodiment 2: Correlation test of detection reagent

[0045] Use the inventive reagent of this method (the specific formula is the same as that in Example 1) and the MB latex enhanced reagent of the contrast reagent A company, and use the automatic 7170 automatic biochemical analyzer to conduct simultaneous tests on 50 human sera (including normal and abnormal samples) according to their respective parameters. Determination and correlation analysis of the measured values. Determination is carried out according to the parameters in the above "MB determination method", and the determination results are shown in figure 2 , X and Y axes are measured values ​​(MB content ng / ml),

[0046] Depend on figure 2 The results show that the correlation between the two reagents is R 2 =0.9895, the regression equation is y=0.9867x+8.6999. The results show that this reagent has a good correlation with imported reagents in the determination of patient serum, and has good specificit...

Embodiment 3

[0048] Embodiment 3: minimum detection limit test

[0049] The purpose of this experiment is to detect the minimum detection sensitivity of the reagent when testing clinical samples.

[0050] Use experimental example 1 reagent, control reagent, standard, blank solution (normal saline and purified water), normal human serum samples, and low-value samples (samples whose values ​​are within ±1 / 3 of the lower limit of the linear range of the reagent).

[0051] Machine: Hitachi 7170 automatic biochemical analyzer.

[0052] Operation steps: Use physiological saline or deionized water to dissolve low-value samples, then dilute 50% to 5 points, test each sample 5 times with the zero point, calculate the average value, and obtain the SD value.

[0053] Result analysis: According to the test data, calculate the SD value and CV value, respectively calculate 1SD, 2SD, starting from the smallest, the value of the average - 2SD is above the zero point average value + 2SD is the minimum det...

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Abstract

The invention discloses a detection kit for measuring the content of myohemoglobin in serum, relating to the field of in-vitro diagnosis in medical immunology. The kit consists of a reagent R1, a reagent R2 and a calibrator, wherein the reagent R1 is a phosphate buffer system; the composition of the reagent R2 is a sensitization latex solution of a crosslinked myohemoglobin antibody; and in the calibrator, the recombinant myohemoglobin is dissolved in the phosphate buffer, and at the same time, glycerin, sucrose, BSA (bovine serum albumin), mannitol and sorbitol are added as protective agents, and sodium azide is used as a preservative. The detection kit disclosed by the invention is simple and convenient to operate, has high sensitivity and good specificity, and can quickly obtain a measurement result; and the measurement result has high accuracy.

Description

technical field [0001] The invention relates to the field of in vitro diagnosis of medical immunity, in particular to a detection kit for measuring the content of myoglobin in serum. Background technique [0002] Acute myocardial infarction (AMI) is one of the diseases with high morbidity and mortality. Currently, the main biochemical markers for AMI detection include creatine kinase isoenzyme (CK-MB), lactate dehydrogenase (LDL), cardiac troponin T (cTnT), and myoglobin (MB). However, the sensitivity of CK-MB, LDL, and cTnT in diagnosing myocardial infarction is not as high as that of MB, which is so far recognized as an early marker of myocardial injury. MB is a binding protein composed of a peptide chain and a heme prosthetic group. It is the main protein that composes skeletal muscle and cardiac muscle, with a molecular weight of 17.8KD. Due to its small molecular weight, it can be quickly released from the ischemic myocardial tissue into the blood. The blood concentra...

Claims

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Application Information

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IPC IPC(8): G01N33/72G01N21/31
Inventor 陈瑛李伟奇房君江张秀文李刚林清玉
Owner 上海睿康生物科技有限公司
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