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Improved cystatin C detection kit

A technology for detecting kits and cystatin, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of large amount of coating and antibody usage, lack of enzymatic amplification reaction, and increased cost of kits, etc. To achieve the effect of shortening the reaction time, increasing the sensitivity and convenient detection

Active Publication Date: 2015-02-18
ZYBIO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, due to the lack of enzymatic amplification reaction in the detection principle of immunoturbidimetric method, the existing cystatin C immunoturbidimetric detection kits do not avoid the disadvantages of low sensitivity and so on.
At the same time, in the process of kit preparation, in order to ensure that the product reaches the required sensitivity, compared with ELISA and chemiluminescence methods, it has higher requirements on the quality of the antibodies used, and the amount of coating and antibody usage is larger, increasing the the cost of the kit
Moreover, mouse monoclonal antibodies, rabbit polyclonal antibodies or goat polyclonal antibodies, which are widely used at present, will inevitably be interfered by rheumatoid factor and heterophilic antibodies during use due to their own properties, which will affect the final detection results.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The kit of the present invention is exemplified as a double reagent, wherein:

[0047] Reagent R1:

[0048]

[0049] Reagent R2:

[0050]

[0051]

[0052] Coated with polystyrene latex particles sensitized by chicken anti-cystatin C yolk antibody, the particle diameter is 100nm, and the concentration is 1%.

[0053] Calibrator:

[0054]

[0055] According to the required concentration of the cystatin C reference calibrator, the corresponding cystatin C standard was added to the above buffer respectively to prepare a set of calibrator with different concentrations.

Embodiment 2

[0056] Embodiment 2: Kit usage method.

[0057] 1. Reagent preparation, the reagent is a liquid double reagent, ready to use after opening the bottle, of which:

[0058] (1) Preparation of reagent R1:

[0059] Configuration buffer 1: stachyose 0.8-3g / L, alum 0.1-1g / L, sodium fructose diphosphate 0.8-3g / L sodium hexametaphosphate 0.05-0.5g / L, glycine 1.5-2.25g / L, solvent For water, PH7.4.

[0060] Add PEG8000 2.7g / L, sodium azide 1.6g / L, bovine serum albumin 2.2g / L, edetate disodium 4.5g / L, sodium chloride 2.5g / L in buffer 1, stir Mix evenly to obtain reagent R1;

[0061] (2) Preparation of reagent R2:

[0062] Step 1: Dialyze the Cystatin C antibody at 4°C, then dilute the Cystatin C antibody to 2 mg / ml with buffer 2 to obtain the Cystatin C antibody dilution; centrifuge the polystyrene latex microspheres with distilled water , wash 3 times;

[0063] Step 2: Dilute the polystyrene latex microspheres washed in step 1 to a mass concentration of 1% with buffer solution 2 (t...

Embodiment 3

[0083] Preparation and use of the comparison kit:

[0084] The buffer solution in the reagent R1 adopts tromethamine buffer solution 75mmol / L, pH7.2, and other reagents and experimental methods are the same as in Examples 1 and 2.

[0085] Reagent R1:

[0086]

[0087] Reagent R2:

[0088]

[0089] Coated with polystyrene latex particles sensitized by chicken anti-cystatin C yolk antibody, the particle diameter is 100nm, and the concentration is 1%.

[0090] Calibrator:

[0091]

[0092] According to the required concentration of the cystatin C reference calibrator, the corresponding cystatin C standard was added to the above buffer respectively to prepare a set of calibrator with different concentrations.

[0093] The performance test of the cystatin C detection kit prepared in Example 3 is the same as in Example 2.

[0094] 1) Minimum detection limit: 5% BSA saline solution is used as a blank sample, and the blank sample should not contain the analyte. The dete...

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Abstract

The invention provides a latex enhanced turbidimetric immunoassay kit for quantitatively detecting cystatin C. The kit comprises a reagent R1 and a reagent R2 which are independent from each other, wherein the reagent R1 is mainly prepared from a buffer solution 1, a stabilizing agent 1, a preservative 1, EDTA, accelerator and a protective agent 1; the reagent R2 is mainly prepared from polystyrene latex microspheres for crosslinking a cystatin C antibody, a buffer solution 2, a stabilizing 2, a preservative 2 and a protective agent 2, wherein the polystyrene latex microspheres and the cystatin C antibody are connected in a covalent cross-linking mode. The detection kit has the advantages of low preparation cost, good stability, good data repeatability and high detection sensitivity, is easy to store, and can be widely applied to clinical biochemical analyzers.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to an improved cystatin C detection kit and its preparation method and application. Background technique [0002] The clinical evaluation of the progress and severity of kidney disease generally takes renal function as a reference, and renal function is generally reflected by glomerular filtration rate (GFR). It is the most important indicator of renal function. According to the source of glomerular filtration rate (GFR) markers, it can be divided into exogenous and endogenous. Exogenous markers include inulin, iohexol, 51Cr-EDTA, 99mTc-DTPA and the like. Endogenous markers include serum creatinine (Scr), urea (Urea), β 2 - Microglobulin (β 2 -M), β-trace protein (BTP) and serum cystatin C (Cystatin C, Cys C). [0003] Cystatin C (Cystatin C, Cys C), also known as γ2 trace basic protein or post-γ globulin, is a kind of cysteine ​​protease inhibitor protein. The gene encodin...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/531G01N33/54313
Inventor 李元丽
Owner ZYBIO INC
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