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Detection reagent for heart-type fatty acid binding protein and preparation method of detection reagent for heart-type fatty acid binding protein

A fatty acid combination and protein detection technology, which is applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of reduced accuracy and achieve high specificity and high analytical sensitivity

Inactive Publication Date: 2015-02-04
中国人民解放军第三0五医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After 1 hour of thrombolytic therapy, the accuracy decreased to 94%

Method used

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  • Detection reagent for heart-type fatty acid binding protein and preparation method of detection reagent for heart-type fatty acid binding protein
  • Detection reagent for heart-type fatty acid binding protein and preparation method of detection reagent for heart-type fatty acid binding protein
  • Detection reagent for heart-type fatty acid binding protein and preparation method of detection reagent for heart-type fatty acid binding protein

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Experimental program
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Effect test

preparation example Construction

[0061] The preparation method of the heart-type fatty acid binding protein detection reagent is as follows:

[0062] 1), adding the coagulant, BSA and preservative to the buffer solution to prepare reagent one;

[0063] 2), heating and boiling chloroauric acid and trisodium citrate according to a mass ratio of 1:1 to prepare a colloidal gold solution; adding the H-FABP antibody to the colloidal gold solution, and adding stabilizers, buffers and preservatives to prepare Obtain reagent two; Wherein, when configuring chloroauric acid and trisodium citrate solution, use 0.2um filter membrane to filter;

[0064] 3) Reagent 1 and reagent 2 are used in combination according to the volume ratio of 5:1 to obtain the heart-shaped fatty acid binding protein detection reagent.

Embodiment 1

[0067] Preparation of colloidal gold particles (all the following operations need to be completed in a high cleanliness and dust-free space)

[0068] 1) Preparation: All the glass containers used are first washed with detergent and rinsed with running water, then soaked in siliconized reagent overnight, siliconized the inner surface of the metal utensils used, rinsed with distilled water, and set aside;

[0069] 2) Prepare 1% (w / v) chloroauric acid and 1% (w / v) sodium citrate solution, and filter with 0.2um membrane;

[0070] 3) Put a magnetic stirrer in a cleaned 1000ml round bottom flask, and add 1000ml ultrapure water;

[0071] 4) Add a certain volume concentration of 1% (w / v) chloroauric acid, stir at a high speed around 600rpm, and heat until the solution boils, then quickly add a certain volume concentration of 1% sodium citrate solution into the flask and keep heating and vigorously stirred for 10 min, the solution color first turned black, and then gradually turned pu...

Embodiment 2

[0075] Preparation of reagent one

[0076] The formula of reagent one is as follows: Tris-HCl buffer solution, 0.9% NaCl (w / v), 2% BSA (w / v), 0.1% NaN3 (w / v), 0.05% Tween20 (w / v), according to The concentration of the Tris quality control buffer was adjusted according to the amount of HCl used to adjust the pH of the buffer.

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Abstract

The invention relates to a detection reagent for heart-type fatty acid binding protein and a preparation method of the detection reagent for the heart-type fatty acid binding protein. The detection reagent is prepared from a reagent I and a reagent II, wherein the volume ratio of the reagent I to the reagent II is 5 to 1; the reagent I comprises a 20-50mmol / L buffer solution, a 2%w / v coagulant, 2%w / v BSA and a 0.1%-0.5%w / v preservative; the reagent II comprises 0.1%-0.5%w / v anti-human H-FABP antibody-marked colloidal gold particles, a 2%-5%w / v stabilizer, the 20-50mmol / L buffer solution and the 0.1%-0.5%w / v preservative; the diameters of the anti-human H-FABP antibody-marked colloidal gold particles are 60nm-90nm. The detection reagent has the advantages of being high in detection sensitivity, large in linear detection range, easy and rapid in detection, good in operability and the like.

Description

【Technical field】 [0001] The invention relates to a heart-shaped fatty acid binding protein detection reagent and a preparation method thereof, in particular to a homogeneous sol particle type H-FABP immunoassay reagent for detecting H-FABP in serum or plasma, and belongs to the field of in vitro diagnostic reagents for medical devices. 【Background technique】 [0002] Heart-type fatty acid-binding protein (H-FABP), a novel small cytosolic protein enriched in the heart, is highly cardiac-specific (i.e., predominantly expressed in cardiac tissue), but also low in tissues other than the heart. concentration expression. After the occurrence of myocardial ischemic injury, H-FABP can be found in the blood as early as 1-3 hours after the onset of chest pain, peaks at 6-8 hours and the plasma level returns to normal within 24-30 hours. Cardiac fatty acid binding cytoplasmic protein consists of 132 amino acids with a molecular weight of 15 kDa. The heart-type fatty acid binding pro...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531
CPCG01N33/6887
Inventor 王会中
Owner 中国人民解放军第三0五医院
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