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Kit (chemiluminiscence method) for determining anti-ribonucleoprotein 70 antibody IgG and manufacturing method thereof

A technology of ribonucleoprotein and chemiluminescence, which is applied in the field of in vitro diagnostic immunoassay, can solve the problems of narrow detection linear range, low detection sensitivity, and high detection cost, and achieve the effects of increased sensitivity, high detection sensitivity, and wide linear range

Inactive Publication Date: 2017-04-26
SHENZHEN YHLO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0019] The current anti-ribonucleoprotein 70 antibody IgG detection technology has the following disadvantages: high detection cost, low detection sensitivity, narrow detection linear range, low reproducibility, inability to quantify, complicated operation, etc.

Method used

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  • Kit (chemiluminiscence method) for determining anti-ribonucleoprotein 70 antibody IgG and manufacturing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Preparation method of anti-ribonucleoprotein 70 antibody IgG assay kit (chemiluminescent method)

[0030] (1) Preparation of nano-magnetic beads coated with anti-ribonucleoprotein 70 antibody IgG antigen:

[0031] Take 50mg of carboxylated magnetic particles (particle size: 0.05-1um) suspension, magnetically separate to remove the supernatant, resuspend with 0.02 M, pH 5.5MES buffer, add 0.5-2mL of newly prepared 10 mg / mL EDC Aqueous solution, activate the carboxyl group on the surface of magnetic beads, add 3-5mg anti-ribonucleoprotein 70 antibody IgG antigen, suspend at room temperature for 2-10 h, magnetically separate, remove the supernatant, and use 0.1 M Tris pH 8.0 containing 2% BSA The buffer solution was resuspended to 1 mg / mL to obtain anti-ribonucleoprotein 70 antibody IgG antigen-coated magnetic particles, which were divided into 5 mL bottles and stored at 4°C for later use.

[0032] (2) Preparation of mouse anti-human IgG-labeled acridinium este...

Embodiment 2

[0040] Embodiment 2: anti-ribonucleoprotein 70 antibody IgG chemiluminescent immunoassay method:

[0041] The present invention uses a fully automatic chemiluminescence immunoassay analyzer as a detection tool, and the methodological mode of the present invention is an indirect method, that is, the instrument adds 10 uL of the sample to the sample diluted 10 times with the sample diluent, then takes 10 ul, and then adds 50 uL of anti-ribose nucleus Magnetic particles coated with protein 70 antibody IgG antigen were reacted for 10 minutes, then magnetic separation was performed; then 50 uL of test diluent and 50 ul of acridinium ester of mouse anti-human IgG were added in sequence, and after 10 min of reaction, magnetic separation was performed. The reaction mixture was sent to the dark room, and 50uL chemiluminescence pre-excitation solution and 50uL chemiluminescence excitation solution were added in turn for luminescence reaction, and finally the luminescence intensity was re...

Embodiment 3

[0042] Example 3: Performance evaluation of anti-ribonucleoprotein 70 antibody IgG assay kit (chemiluminescence method)

[0043] The detection curve is attached figure 1 .

[0044] Sensitivity detection:

[0045]The sensitivity of the anti-ribonucleoprotein 70 antibody IgG assay kit (chemiluminescence method) was calculated by referring to the recommended experimental protocol in the CLSI EP17-A document, and the obtained sensitivity was 0.34AU / mL.

[0046] Linear detection:

[0047] Perform linear analysis on standard products with concentrations of 12.5 AU / mL, 25 AU / mL, 50 AU / mL, 100 AU / mL, and 200 AUmL, and calculate the linear correlation coefficient, r=0.9997. In addition, the kit against ribonucleoprotein 70 antibody IgG samples were detected in the linear range of 3-165 ng / mL.

[0048] Precision determination:

[0049] Take two anti-ribonucleoprotein 70 antibody IgG samples with a concentration of 30 AU / mL and 150 AU / mL, make 3 parallels for each concentration of e...

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Abstract

The invention discloses a kit (chemiluminiscence method) for determining an anti-ribonucleoprotein 70 antibody IgG. The kit comprises magnetic particles coated by anti-ribonucleoprotein 70 antigen, acridinium ester marked by mouse-anti-human IgG, a sample diluent, a test diluent, an anti-ribonucleoprotein 70 antibody IgG calibration product, pre-exciting liquid and exciting liquid. The invention further discloses a manufacturing method of the kit (chemiluminiscence method) for determining the anti-ribonucleoprotein 70 antibody IgG. Compared with an existing kit, the kit has the advantages of being easy to operate, high in sensitivity and wide in detection range.

Description

technical field [0001] The present invention relates to the field of in vitro diagnostic immunoassay, specifically, the present invention provides an anti-ribonucleoprotein 70 antibody IgG side assay kit (chemiluminescent method) and a preparation method thereof. Background technique [0002] The small nuclear riboprotein complex (U-snRNP) plays an extremely important role in the cleavage of precursor messenger RNA. Most RNP particles in the nucleus are U1-snRNP, which is a complex composed of a small fragment of RNP rich in uridine (U1-RNP) and concentrated protein. Anti-ribonucleoprotein 70 antibody (RNP70) is an antibody against the 70kDa protein in U1-RNP, which can be found in 35% to 45% of patients with systemic lupus erythematosus (SLE) and 95% to 100% of patients with mixed connective tissue disease (Mixed Connectivetissue disease, MCTD) patients, is a diagnostic sign of mixed connective tissue disease. The age of onset ranges from 4 to 80 years old, and most patie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N21/76
CPCG01N33/531G01N21/76
Inventor 王刚徐燕玲李路新肖华淼田甜
Owner SHENZHEN YHLO BIOTECH
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