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Lipoprotein phospholipase A2 detection kit

A detection kit and lipoprotein technology, applied in the field of medical detection, can solve the problems of inability to accurately quantitatively measure, falsely increase the detection value, and use a large amount of antibodies, so as to reduce the amount of antibodies, improve accuracy, and reduce production. low cost effect

Inactive Publication Date: 2016-06-29
HANGZHOU WEIXIN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of these methods cannot be used for accurate quantitative determination, and only latex-enhanced immunoturbidimetric method can be used for accurate quantitative determination. Aggregated particles are formed. Under certain wavelength conditions, the turbidity formed by the aggregated particles has a proportional relationship with the corresponding antigen. Therefore, by measuring the turbidity generated by the aggregates, the content of the antigen to be tested in the sample can be measured.
At present, the common methods of coating anti-lipoprotein phospholipase A2 antibody on latex particles are physical adsorption method or random direction coupling method. Interference of factor (RF) and heterophilic antibodies, IgM, IgG type RF or heterophilic antibodies can bind to the Fc region of antibodies coated on latex, resulting in falsely increased detection values

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The preparation of embodiment 1 lipoprotein phospholipase A2 detection kit

[0022] A. Preparation of reagent R1:

[0023] Accurately weigh 5g PEG8000, 10g sodium chloride, 8g sucrose, 5g Tween 20 and 1g sodium azide; dissolve in 1L of 0.25M ammonium chloride buffer, adjust with 1mol / L hydrochloric acid and 1mol / L sodium hydroxide solution The pH value of the solution is 9.0.

[0024] B. Preparation of reagent R2:

[0025] Anti-lipoprotein phospholipase A2 antibody: mouse monoclonal antibody, titer determined by ELISA method is 1:50000.

[0026] Latex particles: use polystyrene latex particles with a particle size of 150-200nm with carboxyl groups.

[0027] 1. Take 1ml (50mg / ml) latex particles, wash with 0.2M, pH5.0 MES solution (2-morpholineethanesulfonic acid buffer) three times, and disperse;

[0028] 2. Add 0.05ml of 10mg / ml EDAC solution freshly prepared with 0.2M, pH5.0 MES solution, and react at room temperature for 1 hour;

[0029] 3. Add 0.5ml of 100mg / ml...

Embodiment 2

[0038] Embodiment 2 kit performance investigation

[0039] Adopt the kit prepared in embodiment 1 to carry out following test:

[0040] 1. Precision test

[0041] Two human serum samples with different lipoprotein phospholipase A2 contents were used to determine the intra-assay precision of the detection kit of the present invention. Three batches of detection kits were used to measure the same human serum sample 20 times respectively, and the inter-batch relative range of the detection kit of the present invention was calculated. The results showed that the intra-assay precision of the detection kit of the present invention was 4.26% and 1.78% (see Table 1), and the inter-assay relative range was 4.38% (see Table 2).

[0042] Table 1

[0043]

[0044] Table 2

[0045]

[0046]2. Linearity measurement test

[0047] Determination of different lipoprotein phospholipase A2 protein content standards, the concentrations of lipoprotein phospholipase A2 protein standards w...

Embodiment 3

[0054] Embodiment 3 test kit stability investigation

[0055] Under the storage condition of 2-8°C, the same serum sample was measured at 0 month, 12 month and 24 month respectively, each sample was measured 20 times, and the average value was taken (see Table 4 for test results). The results show that the difference in measured values ​​is very small, indicating that the detection kit prepared in Example 1 can be stable for at least 24 months under storage conditions of 2-8 degrees.

[0056] Table 4

[0057]

[0058] Measured value difference = (measured value - measured value at 0 months) / measured value at 0 months × 100%

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Abstract

The invention provides a lipoprotein phospholipase A2 detection kit which comprises reagent R1 and reagent R2, the reagent comprises a buffer solution, a stabilizing agent, electrolyte, a surface active agent and preservative; the reagent R2 comprises polystyrene latex particles coupled to an antibody resisting to the lipoprotein phospholipase A2, a buffer solution, a stabilizing agent, a surface active agent and preservative. A directional coupling method is adopted for the polystyrene latex particles sensitized by the antibody resisting to the lipoprotein phospholipase A2, and the preparation steps include activation of the polystyrene latex particles; oxidation of the antibody resisting to the lipoprotein phospholipase A2; coupling of the antibody resisting to the lipoprotein phospholipase A2 to the polystyrene latex particles. The kit is high in detection sensitivity, wide in detection range, good in accuracy and high in interference resistance.

Description

technical field [0001] The invention belongs to the field of medical detection and relates to a detection kit for lipoprotein phospholipase A2. Background technique [0002] Lipoprotein-associated phospholipase A2 (1ipoprotein-associated phospholipase A2, Lp-PLA2) is a phospholipase A2 superfamily closely related to arteriosclerosis and ischemic cardiovascular and cerebrovascular diseases that has attracted widespread attention in recent years, and a new inflammatory marker and an independent risk factor. Its molecular weight is 45KD, synthesized and secreted by mature macrophages and lymphocytes, and regulated by inflammatory mediators. Lipoprotein phospholipase A2 has a pro-inflammatory effect, so the production and release of Lp-PLA2 from inflammatory cells can also be interpreted as an excellent indicator of pro-inflammatory response. Lp-PLA2 produces oxidized molecules in blood vessel walls that are more prone to atherosclerosis and unstable plaques. Elevated levels ...

Claims

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Application Information

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IPC IPC(8): G01N33/573
CPCG01N33/573G01N2333/916
Inventor 石春林商忆文
Owner HANGZHOU WEIXIN BIOTECH CO LTD
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