Latex enhanced turbidimetric immunoassay kit for detection of lipoprotein (a) and preparation method thereof

An immunoturbidimetric and latex-enhanced technology, applied in biological testing, material testing, etc., can solve the problems of poor repeatability of ELISA, low degree of automation, and poor quantitative accuracy, and achieve easy automation, automation, and good stability. Effect

Inactive Publication Date: 2018-01-19
QINGDAO BIOMEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, detection methods for lipoprotein (a) include radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), and latex particle-enhanced turbidimetry. Although RIA has high sensitivity, it is unstable and has poorer repeatability than ELISA. Moreover, there is a risk of radioactive contamination; although the ELISA method has been used clinically for nearly two decades, it still has some fatal shortcomings, such as poor quantitative accuracy, long operation time, and low degree of automation. Generally, it can only be used for qualitative detection

Method used

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  • Latex enhanced turbidimetric immunoassay kit for detection of lipoprotein (a) and preparation method thereof
  • Latex enhanced turbidimetric immunoassay kit for detection of lipoprotein (a) and preparation method thereof
  • Latex enhanced turbidimetric immunoassay kit for detection of lipoprotein (a) and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Preparation and use of lipoprotein (a) kit

[0029] 1. Preparation of lipoprotein (a) antibody latex microspheres: Dilute polystyrene microspheres with a particle size of 30-200 nm with 50 mmol / L MES (pH5.8) buffer solution to form contained polystyrene microspheres The mass concentration is 1%-5% solution; In the latex microsphere solution, add 0.5mg EDAC in every milliliter, stir 30min at room temperature; Add equal volume lipoprotein (a) antibody, stir and couple at room temperature overnight; Centrifuge at a speed of 15000rpm / min for 40 minutes, discard the supernatant; place the precipitate in BSA solution (1%), and suspend it in an ice bath; stir and seal at room temperature for 30 minutes; centrifuge at a speed of 15000rpm / min for 40 minutes, Discard the supernatant; place the precipitate in 50mmol / L Tris (pH7.5) buffer, and suspend it in an ice bath; centrifuge at a speed of 15000rpm / min for 40 minutes, discard the supernatant; repeat steps 7), 8 ). Place the ...

Embodiment 2

[0049] The lipoprotein (a) detection kit was tested for various performance indicators on the Hitachi 7600 analyzer.

[0050] 1. Analytical Sensitivity Evaluation

[0051] Use 5% bovine serum albumin solution as a blank sample, and the blank sample should not contain the analytes. 20 consecutive detections were performed on the biochemical analyzer, and the mean and standard deviation SD of the 20 results were calculated. The detection limit of the reporting method is the mean value of the blank plus two standard deviations (+2SD).

[0052] Measurement times

Mean (mg / L)

SD

X+2SD

20

5.13

0.31

5.93

[0053] As can be seen from Table 1, the sensitivity of the kit of the present invention is 5.93mg / L.

[0054] 2. Linear Experiment Evaluation

[0055] Dilute 1 part of high-value serum (1000mg / L) with pure water at 1:7, 1:3, 1:1, 3:1, 4:0 (samples are numbered 1-5) to prepare 5 samples Samples with different concentrations were tested...

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Abstract

The invention discloses a latex enhanced turbidimetric immunoassay kit for detection of lipoprotein (a). The kit comprises reagents R1 and R2. The reagent R1 contains a Tris buffer solution, sodium chloride, polyethylene glycol and alkylphenol polyoxyethylene. The reagent R2 contains a Tris buffer solution, anti-human lipoprotein (a) antibody coated latex, EDTA, trehalose and alkylphenol polyoxyethylene. By proportionally mixing a sample to be detected and the reagent for a reaction, detecting absorbance change rate at dominant wavelength of 600 nm through a fully automatic biochemical analyzer and referring to absorbance change rate of a calibrator, concentration of lipoprotein (a) in the sample is calculated. The detection method of lipoprotein (a) in the invention is a latex enhanced turbidimetric immunoassay method. The kit of the method has advantages of high detection sensitivity, high accuracy, good stability, strong anti-interference performance and the like.

Description

technical field [0001] The invention relates to an immune turbidimetric detection method, in particular to a kit for a latex-enhanced immune turbidimetric method for detecting lipoprotein (a) and a preparation method thereof. Background technique [0002] Lipoprotein (a) is a special independent plasma lipoprotein, which was discovered by Norwegian geneticist Berg in 1963 when he was studying the genetic variation of low-density lipoprotein. In the late 1980s, it was discovered that lipoprotein (a) was associated with atherosclerosis. [0003] Lipoprotein (a) is a protein in human serum with a structure similar to LDL (low-density lipoprotein), and its density is between HDL (high-density lipoprotein) and LDL. The core of lipoprotein (a) is neutral lipid and APOB100 molecules, surrounded by hydrophilic APOA, which are covalently linked by disulfide bonds; APOA is a characteristic glycoprotein of lipoprotein (a) The composition is mainly composed of a characteristic structu...

Claims

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Application Information

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IPC IPC(8): G01N33/92
Inventor 李娇娇路克国周玭
Owner QINGDAO BIOMEDICAL TECH
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