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Anti-cyclic citrullinated peptide (CCP) antibody detection kit

A technology of anti-cyclic citrullinated peptide and detection kit, which is applied in biological testing, measuring devices, material inspection products, etc., and can solve the problems of not being suitable for laboratories, poor anti-interference ability, inability to distinguish non-specific recognition by molecular weight, etc. , to achieve the effect of improving the detection sensitivity and increasing the change intensity

Active Publication Date: 2014-12-10
上海睿康生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] (1) Non-specific recognition cannot be distinguished according to the size of the molecular weight when analyzing the results;
[0008] (2) The operation is relatively complicated and requires expensive fluorescence microscopes, which are difficult to popularize in many primary hospitals and are not suitable for laboratories with a large number of specimens;
[0009] (3) The background in the fluorescence measurement is relatively high, and it is difficult to use the fluorescence immunoassay technique for quantitative determination;
[0010] (4) Experienced professionals are required to determine the results, and the objectivity of the analysis results is insufficient;
[0011] (5) Only qualitative detection can be carried out, and quantitative measurement cannot be carried out
[0016] (1) Only qualitative and semi-quantitative analysis can be carried out, and the specific amount of the tested substance cannot be obtained
[0017] (2) The operation steps are cumbersome and the test takes a long time
[0018] (3) The detection sensitivity needs to be improved
Therefore, the latex immunoturbidimetric method has great research value, but the latex-enhanced immunoturbidimetric assay kits currently on the market have poor anti-interference ability, low specificity, poor reagent stability, too high reagent background or detection Problems such as narrow linear range need to be improved

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] The CCP detection kit of the present embodiment comprises reagent R1, reagent R2, standard substance and standard substance diluent; Its preparation is specifically as follows:

[0067] Reagent R1:

[0068]

[0069] Various ingredients can be added sequentially at room temperature, sealed and stored for later use.

[0070] Reagent R2:

[0071] The preparation of the latex reagent of the CCP antigen includes the following steps.

[0072] Step 1: activating the anti-CCP antigen in a 20mM MES solution with a pH value of 5.0-5.5;

[0073] Step 2: Adjust the pH of the solution obtained in step 1 to 6.1 with PBS solution, add carboxylated modified polystyrene latex microsphere solution with a particle size of 100-300 nm, and incubate at 37°C for 2-3 hours;

[0074] Step 3: Block the solution obtained in Step 2 with 50 mM Tris with a pH value of 7.0 to 7.5 at 37° C. for more than 5 hours;

[0075] Step 4: after centrifuging or filtering the solution obtained in step 3, ...

Embodiment 2

[0082] The CCP detection kit described in Example 1 is applicable to various types of automatic biochemical analyzers. Taking Hitachi 7170 automatic biochemical analyzer as an example, its operation is shown in Table 1. Analysis method: two-point end-point method, that is, the dosage of reagents R1 and R2 is 150ul and 50ul respectively, and the sample volume is 5ul.

[0083] Adopt this reagent and above-mentioned assay method, adopt the curve of the CCP standard substance (self-made) of 2 kinds of different contents that Hitachi 7170 biochemical analyzer records (such as figure 1 Shown), each point represents a content of the reference standard, wherein the X-axis represents the CCP content (U / ml); Y-axis represents the absorbance.

[0084] Table 1

[0085]

Embodiment 3

[0086] Embodiment 3, linear range

[0087] The high-concentration sample (100.26U / ml) of CCP close to the upper limit of the linear range was diluted with CCP diluent by 1 / 2, 1 / 10, 1 / 15, 1 / 30, and 1 / 60, and a total of 6 Each concentration is detected with the method described in Example 2, and each concentration is repeatedly measured 3 times, and the mean value of the measured concentration and the theoretical concentration are carried out to linear regression analysis, and the calculation regression equation is y=1.0327x+0.5524, and the relevant The coefficient is R 2 =0.9981, showing that the correlation of the kit of the present invention is better in the linear range of 5.00U / ml~100.00U / ml, see figure 2 .

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Abstract

The invention discloses an anti-cyclic citrullinated peptide (CCP) antibody detection kit. The kit comprises a reagent R1, a reagent R2, a standard substance and a standard diluent, wherein the reagent R1 is prepared by washing CCP antigen latex particles by 50mM Tris buffer solution containing 20-500mmol / L first stabilizer and 0.1-1% preservative and then dispersing; and CCP antigens and polystyrene latex microspheres are subjected to chemical crosslinking and then are closed, so that the CCP antigen latex particles are formed. For the kit, the latex-enhanced immunoturbidimetry is adopted for detecting the content of an anti-CCP antibody, the kit has the high sensitivity, the high stability and the high detection speed (the time from determination to result acquisition is only within 5-10min), can detect mass samples on a conventional biochemical analyzer and can greatly improve the detection efficiency.

Description

technical field [0001] The invention relates to latex-enhanced immune turbidimetry technology, in particular to an anti-cyclic citrullinated peptide antibody detection kit. Background technique [0002] Anti-cyclic citrullinated peptide antibody is a kind of rheumatic autoantibody system. Rheumatoid arthritis (RA) has a highly specific citrulline-related autoantibody system, including AKA, APF, AFA, and anti-CCP antibodies. The diagnosis of RA has good specificity. Rheumatoid arthritis (RA) is a common systemic autoimmune disease. The main clinical features are joint synovial inflammation and symmetrical and destructive joint lesions. , easy to repeat, causing great pain to the patient. The incidence rate of this disease in my country is about 0.4%, and it is 0.5~1.0% in the whole world. [0003] Anti-cyclic citrullinated peptide antibody (CCP) is a polypeptide fragment of cyclic filaggrin, an IgG-based antibody, which has good sensitivity and specificity for rheumatoid ar...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/544G01N21/64
CPCG01N33/545G01N33/68
Inventor 李伟奇赵小凤房君江张秀文林清玉
Owner 上海睿康生物科技有限公司
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