Lipoprotein (a) detection kit

A technology for detection kits and lipoproteins, which is applied in biological tests, material inspection products, etc., can solve the problems of easy interference in detection values, false positives in measurement values, and long detection time, so as to avoid false positives in measurement values ​​and be accurate. High performance and low production cost

Active Publication Date: 2013-06-12
NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, known methods for detecting the concentration of lipoprotein (a) include enzyme-linked immunoassay, radioimmunoassay, fluorescent immunoassay and other detection methods. The operation of the above-mentioned determination methods is relatively complicated, the detection takes a long time, and the shortcomings of excessive waste of resources , not suitable for routine testing
A lipoprotein (a) detection kit is disclosed on the website of the State Intellectual Property Office of China. It adopts latex-enhanced immunoturbidimetry, which has the advantages of good stability and convenient detection. However, the rheumatoid factor contained in human serum is harmful to the The detection value of the kit is prone to interference, which makes the measured value of individual serum with high content of rheumatoid factor appear false positive
In addition, the existing latex-enhanced immunoturbidimetric method uses physical adsorption to label polyclonal antibodies on polystyrene latex particles without carboxyl groups. The polystyrene latex particles without carboxyl groups have poor stability and physical The effect of adsorption is also not good, it is easy to cause the antibody to fall off from the surface of latex particles

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Experimental program
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Embodiment 1

[0025] Preparation and determination method of lipoprotein (a) detection kit

[0026] The kit of the present invention relates to the main raw materials of reagents as follows:

[0027] 1. The goat anti-human apolipoprotein (a) polyclonal antibody made by our company according to the conventional method, after affinity purification, the titer is 1:100000 by indirect ELISA method, which meets the requirements of this reagent. The antibody has good specificity and no cross-reaction with other antigens.

[0028] 2. In the present invention, only polystyrene latex particles with carboxyl groups are used for experiments, and the corresponding detection wavelength is 600nm.

[0029] 3. Recombinant apolipoprotein (a) is self-made by our company, expressed in Escherichia coli or yeast, and purified by affinity and ion exchange. The apolipoprotein (a) has high purity and good stability, and is used as the standard product of this reagent .

[0030] The main reagents and reference ...

Embodiment 2

[0036] Lipoprotein (a) detection kit was tested on various performance indicators on Hitachi 7060

[0037] 1. High value linear determination

[0038] The recombinant apolipoprotein (a) was added to the blank serum, and the final concentration of the apolipoprotein (a) in the serum was 1000 mg / l. Serum was diluted with 0.9% NaCl to obtain 5 serum samples containing different concentrations of apolipoprotein (a). Theoretical concentration of apolipoprotein (a) is 62.5mg / L, 125mg / L, 250mg / L, 500mg / L, 1000mg / L respectively. The blank serum was used as a sample with apolipoprotein (a) concentration of 0. Each sample is measured 3 times with the lipoprotein (a) kit among the present invention, and average value is obtained. Correlation analysis is carried out to measured value, as can be seen from Table 1, the highest detection range of the detection kit of the present invention can reach 1000mg / L, see figure 2 , Y-axis measured value, X-axis theoretical value (apolipoprotein ...

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Abstract

The invention relates to a lipoprotein (a) detection kit which comprises a reagent R1, a reagent R2 and a reference substance, wherein the reagent R1 is an HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) buffer system which comprises 0.5-10 g / L HEPES, 2-20 g / L sodium chloride, 0.05-1.0 ml / L Tween-20, 0.1-2 g / L bovine serum albumin, 5-25 g / L polyethyleneglycol-6000, 1-5 g / L EDTA (ethylene diamine tetraacetic acid) and 0.1-2 g / L Proclin 300; the reagent R2 comprises a polystyrene latex particle mixture which is prepared by a chemical crosslinking method, coated with an anti-human-apolipoprotein (a) polyclonal antibody and provided with carboxylic groups on the surface, a 0.5-10 g / L HEPES buffer solution and aspartame, wherein the weight-to-volume ratio of the aspartame to the reagent R2 is (0.01-0.5):100 (g / L); and the reference substance is a human serum or serum-matrix-like liquid with human recombinant apolipoprotein (a). The kit provided by the invention has the advantages of high detection sensitivity, high accuracy, favorable precision, favorable linearity within detection range, high stability, low production cost and low blank absorbance, and has anti-interference performance.

Description

technical field [0001] The invention relates to a medical detection kit, in particular to a kit for measuring lipoprotein (a) in human serum by using latex-enhanced immune turbidimetry. Background technique [0002] Lipoprotein (a) (Lipoprotein (a), LP (a)) is a lipoprotein similar to low-density lipoprotein, and its core part contains triglycerides, phospholipids, and cholesterol similar to low-density lipoprotein components , cholesteryl ester and other lipids and apolipoprotein B100 (ApoB100), also contains apolipoprotein (a) (Apolipoprotein (a), Apo (a)) which is not found in low-density lipoprotein. Therefore, the content of lipoprotein (a) in serum is usually reflected by detecting the concentration of apolipoprotein (a) in medicine. The cDNA sequence of apolipoprotein (a) was reported in 1993, and the primary structure of the protein was deduced, and its relative molecular mass was found to be (250-800kDa), containing 4529 amino acids. Apolipoprotein (a) is linked t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/92
Inventor 邹炳德邹继华方亮刘献文周海滨
Owner NINGBO MEDICAL SYSTEM BIOTECHNOLOGY CO LTD
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