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Chemiluminescent enzyme-linked immunosorbent assay kit for detecting lipoprotein-associated phospholipase a2 and preparation method thereof

A chemiluminescence enzyme and immunoreagent technology, which is applied in the detection field of chemiluminescence enzyme-linked immunosorbent technology, can solve the problem of not seeing human Lp-PLA2 kits, etc., and achieve the effect of increasing detection specificity and enhancing difference.

Active Publication Date: 2015-09-16
同昕生物技术(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, there has been no report on using the correlation between Lp-PLA2 and cardiovascular and cerebrovascular diseases to prepare a kit for detecting the level of human Lp-PLA2

Method used

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  • Chemiluminescent enzyme-linked immunosorbent assay kit for detecting lipoprotein-associated phospholipase a2 and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. Preparation of Chemiluminescent ELISA Kit for Detection of Lipoprotein-Associated Phospholipase A2 and Its Detection Method

[0029] 1. Preparation of chemiluminescent ELISA kit for detection of lipoprotein-associated phospholipase A2

[0030] 1. Preparation of sample treatment solution

[0031] Sample treatment solution formula: composed of A solution and B solution, wherein A solution is 7.5g ammonium chloride, 1.2g casein, 1.2g sodium stearate, add purified water to make up to 1L, that is, A solution contains 7.5g / L ammonium chloride, 1.2g / L casein and 1.2g / L sodium stearate; B solution is 0.5M Tris-HCl solution (PH7.6).

[0032] 2. Coating anti-Lp-PLA2 monoclonal antibody as a chemiluminescence reaction plate

[0033] The formula of the following antibody coating solution is:

[0034] 0.36g NaH 2 PO 4 ·H 2 O, 3.10 g Na 2 HPO 4 2H 2 O, dilute to 1L, pH7.6.

[0035] Milky white opaque polystyrene 96-well chemiluminescent microtiter plate (Porvair...

Embodiment 2

[0069] Embodiment 2, the test kit that contains sample treatment solution is compared with the test kit that does not contain sample treatment solution

[0070] Experimental group: samples were processed by the sample treatment solution of the present invention

[0071] Control group: samples were not treated.

[0072] The comparative experiment was carried out with the above-mentioned experimental group and control group. 100 samples of normal control and 100 samples of cardiovascular and cerebrovascular diseases were tested in parallel by the experimental group and the control group. The detection method was referred to the above-mentioned Example 1. The detection procedures and results were judged in strict accordance with the instructions of each reagent. The test results are shown in Table 1, Table 2 and Table 3:

[0073] Table 1 Test results of the control group kit

[0074] group

Number of cases

+

-

Positive rate (%)

normal control gro...

Embodiment 3

[0080] Embodiment 3, kit sensitivity and accuracy test

[0081] 1. Kit sensitivity experiment

[0082] 1. Carry out 20 tests on the zero standard solution, and the average value of the test results plus 3 times the standard deviation is used as the minimum detection limit of the kit.

[0083] Table 4 Zero standard measurement results statistical table ng / mL

[0084] sample number

1

2

3

4

5

6

7

8

measured value

0.06

0.07

0.08

0.06

0.07

0.05

0.06

0.05

sample number

9

10

11

12

13

14

15

16

measured value

0.04

0.05

0.08

0.08

0.06

0.06

0.06

0.07

sample number

17

18

19

20

average

standard deviation

Minimum detection limit

measured value

0.06

0.07

0.04

0.06

0.06

0.01

0.1

[0085] It can be seen from Table 4 that the minimum detection limi...

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Abstract

The invention discloses a chemiluminescence enzyme-linked immunosorbent assay kit for detecting lipoprotein-related phospholipase A2 and a preparation method of the assay kit. The chemiluminescence enzyme-linked immunosorbent assay kit for detecting lipoprotein-related phospholipase A2, which is disclosed by the invention, comprises a chemical luminous panel enveloped by a monoclonal antibody for resisting the protein-related phospholipase A2. The chemiluminescence enzyme-linked immunosorbent assay kit is characterized by also comprising a sample treating solution, wherein the sample treating solution mainly comprises a solution A and a solution B; the solution A contains 7.5g / L ammonium chloride, 1.2g / L casein and 1.2g / L sodium stearate; the solution B contains 0.5M Tris-Hcl; PH is 7.6. In order to reduce Lp-PLA2 luminous detection noise to the maximal extent, the kit disclosed by the invention also comprises a unique blood sample treatment agent formula, so that quenching of a non-specific light source is facilitated, the difference between detecting light and background noise light is greatly enhanced, and the detection specificity is increased.

Description

technical field [0001] The invention relates to the detection field of chemiluminescent ELISA technology, in particular to a chemiluminescent ELISA kit for detecting lipoprotein-related phospholipase A2 and a preparation method thereof. Background technique [0002] Lipoprotein-associated phospholipase A2 (Lp-PLA2) is a calcium ion-independent phospholipase in the phospholipase A2 superfamily. It was originally found to degrade platelet-activating factor and was once called platelet-activating factor acetylhydrolase. It was cloned for the first time in 1995. Its coding gene (PLA2G7) has 12 exons and its chromosome is located at 6p21.2212. It is a serine lipase composed of 441 amino acid residues and its relative molecular mass is 50×10 3 (50kDa). Lp-PLA2 uses biomembrane phospholipids as a natural substrate to preferentially act on the ester bond in the short fatty acyl chain on the sn22 position of oxidized phospholipids, making the highly water-soluble phospholipids produ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/573G01N33/92G01N2333/916
Inventor 吴凡焦守恕
Owner 同昕生物技术(北京)有限公司
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