Immunoturbidimetric assay apolipoprotein E detection kit

A technology for detecting kits and apolipoproteins, which is applied in the field of kits and can solve the problems of direct application to an automatic biochemical analyzer, inconvenient operation and poor anti-interference ability.

Inactive Publication Date: 2013-11-20
上海睿康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Known methods for measuring ApoE include chromatography, electrophoresis, and immunoassay. Among them, chromatography and electrophoresis are cumbersome to operate, and they cannot perform batch sample analysis and directly go to full-automatic biochemical analyzers. Immunodiffusion, radioimmunoassay, fluorescence-labeled immunoassay, enzyme-labeled immunoassay, and chemiluminescence immunoassay also have many deficiencies; if special equipment is required, the sample needs to be processed, and it cannot be used on a fully automatic biochemical analyzer Perform batch monitoring and analysis, etc.
The immunoturbidimetric method commonly used in clinical practice is popular because of its small amount of sample, which can be directly analyzed in batches on a fully automatic biochemical analyzer, and is easy to operate; however, the currently established methods and reagents have some shortcomings and deficiencies, mainly in : First, the antibody titer is not high, and the specificity, affinity, and affinity are not good; second, the calibration serum is based on human serum, although the human serum with negative hepatitis B surface an

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The composition of the assay kit is as follows:

[0029] The main raw materials needed for detection kit and standard of the present invention are as follows:

[0030] Anti-human apolipoprotein E antiserum was purchased from Mildland Company, and ApoE antigen (purchased from Mildland Company) was used to prepare the standard and quality control materials required for this reagent.

[0031] The R1 reagent was prepared as follows:

[0032] Reagent R1:

[0033] PH7.4 Tris buffer 45mmol / L

[0034] Polyethylene glycol 8000 100mmol / L

[0035] Disodium EDTA 30mmol / L

[0036] NaN3 9‰

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

[0038] Reagent R2:

[0039] PH7.4 Tris buffer 45mmol / L

[0040] Polyethylene glycol 8000 100mmol / L

[0041] Disodium EDTA 30mmol / L

[0042] Anti-human apolipoprotein E antiserum 40%

[0043] NaN3 9‰

[0044] Bovine serum matrix calibrator pr...

Embodiment 2

[0049] Correlation test of detection reagents:

[0050] 1. Correlation test:

[0051] Using the inventive reagent of this method (the specific formula is the same as in Example 1) and the ApoE immunoturbidimetric reagent of A company as a control reagent, 50 human sera (including normal and abnormal samples) were simultaneously analyzed according to their respective parameters by using an automatic 7170 automatic biochemical analyzer. Measurements were carried out, and correlation analysis was carried out on the measured values. According to the parameters in the above-mentioned "ApoE assay method", the assay results are shown in figure 2 , X and Y axes are measured values ​​(ApoE content mg / dL),

[0052] Depend on figure 2 The results show that the correlation between the two reagents is R 2 =0.9898, the regression equation is y = 0.9856x + 1.4783. The results show that this reagent has a good correlation with imported reagents in the determination of patient serum, and ...

Embodiment 3

[0055] Minimum detection limit test:

[0056] The purpose of this implementation is to detect the minimum detection sensitivity of the reagent when testing clinical samples. Use the reagents of Example 1, control reagents, standard products, blank solutions (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).

[0057] Machine: Hitachi 7170 automatic biochemical analyzer.

[0058] 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.

[0059] Result analysis: According to the detection 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 detection ...

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Abstract

The invention relates to an immunoturbidimetric assay apolipoprotein E detection kit, and relates to the technical field of kit. The kit is composed of three parts which are an R1 reagent, an R2 reagent, and a calibrator. The R1 reagent is composed of 40-50mmol/L of a buffer solution with a pH value of 7, 70-150mmol/L of a surfactant, 15-30mmol/L of disodium ethylenediamine tetraacetate, and 0.1-1% of a preservative. The R2 reagent is prepared by adding anti-human-apolipoprotein-E antiserum with a volume ratio of 30-50% into a certain amount of the R1 reagent. The calibrator is 80-200g/L ApoE antigen, and also comprises 0.2-15% of a preservative, 1-4% of a stabilizing agent, and 1-10% of a surfactant, wherein the percentages are human serum matrix volume amount percentages. The kit has the advantages of simple operation, high sensitivity, good specificity, fast determination, and accurate and reliable result.

Description

technical field [0001] The invention relates to the technical field of kits, in particular to an immunoturbidimetric method apolipoprotein E detection kit. Background technique [0002] Apolipoprotein E (ApoE) is a glycoprotein of 299 amino acids bound to phospholipids. Its molecular weight is 34kD. ApoE can be synthesized in various tissues, but the liver is the main one. First, a propeptide containing 317 amino acids is synthesized. Mature ApoE is formed by ylation and asialolation of extracellular fluid; ApoE is secreted into the blood and transferred to lipoproteins. ApoE is a structural protein necessary for lipoproteins. It can participate in lipoprotein metabolism, regulate cholesterol levels, activate lecithin sterol lipid transferase (LCAT) by binding to the receptors of various tissue cells, and has a certain immune function. Regulatory effect. Determination of ApoE provides an important theoretical basis for the diagnosis and treatment of hyperlipidemia, athe...

Claims

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Application Information

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IPC IPC(8): G01N33/92G01N21/31
Inventor 赵小凤李伟奇房君江张秀文林清玉
Owner 上海睿康生物科技有限公司
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