Lipoprotein-related phospholipase A2 content detection kit and preparation method thereof

A technology for detecting kits and phospholipases, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of cumbersome operation, low accuracy, and poor stability of detection methods, and achieve good bottle opening stability and guarantee The effect of stability and good specificity

Active Publication Date: 2015-07-15
WUHAN LIFE ORIGIN BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ELISA has excellent characteristics such as high detection accuracy and good sensitivity, but the detection method is cumbersome to operate, takes a long time to detect, and the samples can only be detected in batches, which cannot meet the timeliness, repeatability and operation of the detection indicators of the hospital. Simple and other characteristic requirements; the accuracy of enzymatic detection is not high, which can easily lead to errors in test results, resulting in misjudgment of results and adverse effects on patients
Latex immunoturbidimetric method is also a method for determining Lp-PLA2, which is based on common immunoturbidimetric method, but it overcomes the defect of low sensitivity of ordinary immunoturbidimetric method, and also overcomes the defect of ELISA method. Simple operation, short detection time, high sensitivity, wide linear range and many other excellent characteristics
At present, there are also some Lp-PLA2 latex immunoturbidimetric products and kits, but these kits have problems such as narrow linear range, poor stability, poor detection sensitivity and specificity, etc., and further improvement is needed to enable research and development. And the kits produced can better meet the needs of clinical testing

Method used

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  • Lipoprotein-related phospholipase A2 content detection kit and preparation method thereof
  • Lipoprotein-related phospholipase A2 content detection kit and preparation method thereof
  • Lipoprotein-related phospholipase A2 content detection kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Preparation of latex particle solution coated with lipoprotein-related phospholipase A2 antibody:

[0039] (1) Washing and activation of latex particles: Take 0.18g of 160nm polystyrene latex particles (purchased from PolyMicrospheres) in a 50mL centrifuge tube, and then add 15mL of MES buffer with pH 7.0 to the centrifuge tube, shake and disperse Evenly, put the centrifuge tube in a centrifuge at 25000 rpm, centrifuge for 20 minutes, discard the supernatant, and repeat the washing twice; add 10 mL of MES buffer with pH 7.0 to the washed latex particles to resuspend the latex particles. Accurately weigh 35mg of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and 35mg of N-hydroxysulfosuccinimide (NHS) and dissolve each with 1mL of MES buffer with pH 7.0 After the dissolution is complete, add each solution to the latex solution, shake and mix evenly, place on a constant temperature shaker at 30°C, 200 revolutions / min, and react for 1.5 hours;

[0040] (2) Antibody sensi...

Embodiment 2

[0058] Preparation of latex particle solution coated with lipoprotein-related phospholipase A2 antibody:

[0059] (1) Washing and activation of latex particles: Take 0.1g of 120nm polystyrene latex particles (purchased from PolyMicrospheres) in a 50mL centrifuge tube, then add 10mL of MES buffer with pH 7.0 to the centrifuge tube, and shake Disperse uniformly, place the centrifuge tube in a centrifuge at 25,000 rpm, centrifuge for 30 minutes, discard the supernatant, and repeat the washing twice; add 10 mL of MES buffer with pH 7.0 to the washed latex particles to resuspend the latex particles. Accurately weigh 50mg of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and 50mg of N-hydroxysulfosuccinimide (NHS) with 1mL of pH 7.0 MES buffer respectively Dissolve, add each solution to the latex solution after the dissolution is complete, shake and mix evenly, place on a constant temperature shaker at 30°C, 200 revolutions / min, and react for 2 hours;

[0060] (2) Antibody sensiti...

Embodiment 3

[0078] Preparation of latex particle solution coated with lipoprotein-related phospholipase A2 antibody:

[0079] (1) Washing and activation of latex particles: Take 0.15g of 180nm polystyrene latex particles (purchased from PolyMicrospheres) in a 50mL centrifuge tube, and then add 15mL of MES buffer with pH 7.0 to the centrifuge tube, shake and disperse Evenly, put the centrifuge tube in a centrifuge at 25000 rpm, centrifuge for 20 minutes, discard the supernatant, and repeat the washing twice; add 10 mL of MES buffer with pH 7.0 to the washed latex particles to resuspend the latex particles. Accurately weigh 40mg of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and 40mg of N-hydroxysulfosuccinimide (NHS) with 1mL pH 7.0 MES buffer respectively The solution is dissolved. After the dissolution is complete, add each solution to the latex solution, shake and mix evenly, and place it on a constant temperature shaker at 30°C at 200 revolutions / min for 1.5 hours;

[0080] (2) An...

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Abstract

The invention discloses a lipoprotein-related phospholipase A2 content detection kit and a preparation method thereof, belonging to the field of detection of lipoprotein-related phospholipase A2 content. The kit consists of a reagent I and a reagent II which are independent of each other, wherein the reagent I comprises the following components: a biological buffer agent, a coagulant, a preservative, a chelating agent and water; the reagent II comprises the following components: latex particles coated with a lipoprotein-related phospholipase A2 antibody, a biological buffer agent, a surfactant, a preservative, a suspending agent and water. The latex particles coated with the lipoprotein-related phospholipase A2 antibody are prepared by adopting latex particles with the particle size of 120-180nm, and the stability, sensitivity and wide detection linear range of the kit can be effectively guaranteed. Meanwhile, the latex particles coated with the lipoprotein-related phospholipase A2 antibody are prepared by adopting a specific method, so that the bottle-opening stability and detection specificity of the kit are relatively good, and the product is suitable for popularization and application of clinical detection.

Description

Technical field [0001] The invention belongs to the detection field of lipoprotein-related phospholipase A2 content, and particularly relates to a lipoprotein-related phospholipase A2 content detection kit and a preparation method thereof. Background technique [0002] Lipoprotein-associated Phospholipase A2 (Lipoprotein-associated Phospholipase A2, Lp-PLA2) is encoded by the PLA2G7 gene. Because lipoprotein-associated phospholipase A2 has the activity of degrading platelet activating factor, it is also called degrading platelet activating factor (platelet- activating factor acetylhydro-lase, PAF-AH). Lipoprotein-related phospholipase A2 is a member of the phospholipase A2 superfamily. Unlike other members of the phospholipase A2 family, Lp-PLA2 does not require calcium ions to maintain its catalytic activity. So far, two types of Lp-PLA2 are known: plasma type Lp-PLA2, cell type Lp-PLA2. The plasma type Lp-PLA2 is a protein composed of 441 amino acids with a molecular weight o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/573
CPCG01N33/544G01N33/573G01N33/92G01N2333/916
Inventor 不公告发明人
Owner WUHAN LIFE ORIGIN BIOTECH LTD
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