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Detection kit for diagnosing RA, preparating kit, and method for completing standard of quality detection

A detection kit and detection standard technology, applied in the field of biomedicine, can solve problems such as reduced affinity and influence on experimental results

Inactive Publication Date: 2006-07-05
SHANGHAI KEXIN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using citrulline peptides as antigens has certain disadvantages: First, polystyrene is often used in ELISA detection, and citrulline peptides are often absorbed by polystyrene, which affects the experimental results
Although this peptide contains all the antigenic sites, due to this reason, the affinity of the antibody for the peptide is reduced

Method used

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  • Detection kit for diagnosing RA, preparating kit, and method for completing standard of quality detection
  • Detection kit for diagnosing RA, preparating kit, and method for completing standard of quality detection
  • Detection kit for diagnosing RA, preparating kit, and method for completing standard of quality detection

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Experimental program
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Effect test

preparation example Construction

[0134] ①Preparation of standard products: The concentration unit used for the 5 standard products in this kit is relative unit (RU / ml), and the setting method is as follows:

[0135] OD 450 =1.8~2.0, set the concentration unit as 100RU / ml as the upper limit;

[0136] OD 450 =0.9~1.0, set the concentration unit as 50RU / ml;

[0137] OD 450 =0.4~0.5, set the concentration unit as 25RU / ml;

[0138] OD 450 =0.2~0.3, set the concentration unit as 12RU / ml;

[0139] OD 450 =0.1~0.15, set the concentration unit as 6RU / ml;

[0140] OD 450 =0.06~0.08, set the concentration unit as 3RU / ml;

[0141] OD 450 =0.01~0.06, set the concentration unit as 0RU / ml as the lower limit;

[0142] Draw a standard curve, set 5 measurement points, and prepare standard products No. 1 to No. 5 (0, 6, 25, 50, 100 RU / ml) respectively.

[0143] Preparation of Standard No. 5: Take anti-CCP antibody positive and OD 450 = 1-1.2ml serum of RA patients with 1.8-2.0, dilute with 100ml-120ml sample diluen...

Embodiment 1

[0191] Preparation of controls

[0192] ①Preparation of standard products

[0193] Preparation of Standard No. 5: Take anti-CCP antibody positive and OD 450 = 1ml of serum from RA patients with 1.8-2.0, dilute with 100ml sample diluent, mix thoroughly; take 50ml of it, divide it into standard vials, 1ml per bottle, marked as standard -5, at 2-8°C storage.

[0194] Preparation of No. 4 standard product: Take the remaining 50ml above, add the sample diluent to 50mll, and mix well; then take 50ml of it and pack it into standard vials, 1ml per bottle, and mark it as standard product-No. 4, 2-8 Store at ℃.

[0195] Preparation of No. 3 standard product: take the remaining 50ml above, add the sample diluent to 50mll, and mix well; then take 50ml of it and pack it into standard vials, 1ml per bottle, and mark it as standard product-No. 3, 2-8 Store at ℃.

[0196] Preparation of No. 2 standard product: Take the remaining 50ml of the above, add the sample diluent to 50mll, and mix...

Embodiment 2

[0201] Preparation of liquid reagents

[0202] ①Preparation of concentrated washing liquid: Take the 10 times concentrated washing liquid as an example, the ratio is as follows:

[0203] Sodium chloride (NaCL): 217.5g

[0204] Disodium hydrogen phosphate (Na 2 HPO 4 12H 2 O): 28.75g

[0205] Sodium dihydrogen phosphate (NaH 2 PO 4 2H 2 O): 5g

[0206] Tween-20 (TWEEN-20): 12.5ml

[0207] Purified water: 2.5L

[0208] Preparation method: weigh 217.5g of NaCl, 28.75g of NaCl 2 HPO 4 12H 2 O and 5g NaH 2 PO 4 2H 2 O, dissolved in 2.2L purified water. Adjust the pH value to 7.4 with NaOH, then add 12.5ml of Tween-20, and finally add water to 2.5L. Packed into 50ml / bottle, at 15psi (1.05kg / cm 2 ) was sterilized by high pressure steam for 20 minutes and stored at 2°C.

[0209] ②Preparation of sample dilution buffer, the ratio is as follows:

[0210] Bovine Serum Albumin (BSA for short): 25g

[0211] 1% thimerosal sodium salt: 12.5ml

[0212] Gentamicin (40mg / ml...

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Abstract

The invention discloses a diagnosis reagent box for arthritis pauperum and the preparing method thereof as well as a method of completing a quality detection standard. The reagent box is a reagent box for detecting cyclic citrullinated peptide antibodies. The preparing method comprises coating antigens, preparing a contrast and preparing a liquid reagent. The quality detection standard comprises a finished product batched detection standard and a finished product periodic detection standard. And the reagent box has very high positive forecasting value, simple to operate and high- sensitivity and able to be used in fast detection of large numbers of samples and qualitative and quantitative detection of anti-CCP antibodies.

Description

technical field [0001] The present invention relates to the technical field of biomedicine, in particular to a diagnostic kit for autoimmune diseases, a preparation method thereof and a method for completing quality inspection standards, more specifically to a diagnostic kit for rheumatoid arthritis A diagnostic kit and a method for its preparation and fulfillment of quality testing standards. Background technique [0002] (1) Rheumatic diseases [0003] The human immune system has the function of distinguishing "self" from "non-self", and it will not produce an immune response to its own tissues and cells. However, when the human immune system is abnormal, it may also have an immune response to its own tissues and cells, thereby destroying or injuring its own tissues or cells, that is, autoimmune diseases. One of its important signs is the formation of anti-self tissue components. The examination of autoantibodies is an important diagnostic tool for autoimmune diseases. ...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/532G01N33/53G01N33/68G01N33/48C07K16/00
Inventor 倪健杨子义魏国兰罗鹏
Owner SHANGHAI KEXIN BIOTECH
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