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IV type collagen test kit and test method thereof

A collagen detection kit technology, applied in the biological field, can solve the problems of complex operation, unstable measurement results, expensive instruments, etc., and achieve the effect of high detection sensitivity

Inactive Publication Date: 2016-08-03
BEIJING HOMA BIOLOGICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Radioimmunoassay has problems such as complex operation, unstable measurement results, short storage time of reagents, radioactive contamination, and expensive instruments; time-resolved fluorescent immunoassay needs to be labeled with rare metals; chemiluminescence can only emit light once and is susceptible to environmental interference ;Enzyme-linked immunosorbent method has good specificity and high sensitivity, easy operation, stable reagents, and no pollution threat to the environment

Method used

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  • IV type collagen test kit and test method thereof
  • IV type collagen test kit and test method thereof
  • IV type collagen test kit and test method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0087] Take the preparation of 1L as an example:

[0088] (1) Weigh Tris (tris(trihydroxymethylaminomethane, molecular formula: (HOCH2)3CNH2) 1.56g and NaCl4.23g in a 1L beaker; use a pipette to measure 0.2ml of Proclin-300 in a beaker of a small amount of purified water After it is completely dissolved, pour it into the above 1L container;

[0089] (2) Measure 800ml of purified water in a beaker with a measuring cylinder, stir well until it is completely dissolved; adjust the pH with 4MHCL or 4MNaOH, and measure the range between 7.35-7.45;

[0090] (3) Weigh 0.9 g of bovine gamma globulin IgG in a beaker of 800 ml of purified water;

[0091] (4) The final volume is 1000ml. After it is completely dissolved, measure the pH value. The range is between 7.35 and 7.45, which meets the requirements. Filter with a 0.2um filter;

[0092] Reagent R 2 formulation (Table 1)

[0093]

Embodiment 2

[0095] A type IV collagen (IV-COL) test kit of the present invention comprises a magnetic separation reagent, a reagent R 1 , Reagent R 2 , standard substance, quality control substance, calibrator, cleaning concentrate, diluent and luminescent substrate, among which: magnetic separation reagent: nano-magnetic microspheres labeled with mouse anti-human IV-COL monoclonal antibody, the The concentration of nanomagnetic microspheres labeled with mouse anti-human IV-COL monoclonal antibody is 167 μg / ml; Reagent R 1 : contains alkaline phosphatase-labeled IV-COL antibody, and the concentration of the alkaline phosphatase-labeled IV-COL antibody is 1 μg / ml; reagent R 2 : Buffer containing bovine gamma globulin components; standard, quality control and calibrator: bovine serum albumin component five (BSAV) solution containing a certain amount of IV-COL antigen, the concentration of the standard is 0 (S0), 30(S1), 90(S2), 180(S3), 450(S4), 900(S5) ng / ml, the concentration of the qua...

Embodiment 3

[0186] A type IV collagen (IV-COL) test kit of the present invention comprises a magnetic separation reagent, a reagent R 1 , Reagent R 2 , standard substance, quality control substance, calibrator, cleaning concentrate, diluent and luminescent substrate, among which: magnetic separation reagent: nano-magnetic microspheres labeled with mouse anti-human IV-COL monoclonal antibody, the The concentration of nanomagnetic microspheres labeled with mouse anti-human IV-COL monoclonal antibody was 132 μg / ml; Reagent R1: containing alkaline phosphatase-labeled IV-COL antibody, the alkaline phosphatase-labeled IV-COL COL antibody concentration was 0.6 μg / ml; Reagent R2: buffer containing bovine gamma globulin fraction; standard, quality control and calibrator: bovine serum albumin fraction five (BSAV) containing a certain amount of IV-COL antigen ) solution, the concentration of the standard substance is 0(S0), 50(S1), 100(S2), 250(S3), 500(S4), 1000(S5) ng / ml, the concentration of the...

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Abstract

The invention discloses an IV type collagen test kit which is composed of a magnetic separation reagent, a reagent R1, a reagent R2, a standard product, a quality control product, a calibrating product, cleaning concentrate, diluent and a light-emitting substrate .The invention further discloses a preparation method of the kit .The chemical light-emitting technology and immune magnetic particles are combined through the kit, a reaction system close to a uniform phase is provided, and compared with an existing enzyme linked immunosorbent assay technology, the kit has the advantages that specificity and sensitivity are higher, the time for obtaining a detection result is short, operation modes are simple and convenient, the detection result is accurate and reliable, and product cost is greatly reduced.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for determining the content of type IV collagen in serum and a test method thereof. Background technique [0002] Collagen Type IV (CIV for short) is a network structure formed by molecular cross-linking. It has two terminals. The 7s collagen component is the tetramer of the amino terminal of type IV collagen, and NC1 is the carboxyl group of type IV collagen. The terminal dimers are involved in the formation of oligomers. Type IV collagen is the main component of the liver basement membrane and is present in the hepatic portal vascular area, around the central vein, and along the sinusoidal space. [0003] Type IV collagen is deposited in the extracellular matrix without removing telopeptides during anabolism, so the elevated content of type IV collagen in blood may reflect the accelerated turnover rate of hepatic sinusoidal basement membrane. Both basic and clinical studies...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 朱驰
Owner BEIJING HOMA BIOLOGICAL ENG