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A kind of preparation method of myoglobin detection reagent

A technology for detecting reagents and myoglobin, which is applied in the field of in vitro diagnostic reagents for medical devices, can solve problems such as elevated levels and poor specificity, and achieve high analytical sensitivity and high specificity

Inactive Publication Date: 2016-03-30
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

However, the specificity is poor, and diseases such as skeletal muscle injury, trauma, and renal failure can all lead to an increase in

Method used

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  • A kind of preparation method of myoglobin detection reagent
  • A kind of preparation method of myoglobin detection reagent
  • A kind of preparation method of myoglobin detection reagent

Examples

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Comparison scheme
Effect test

preparation example Construction

[0047] The preparation method of the myoglobin detection reagent is as follows:

[0048] 1), adding the coagulant, BSA and preservative to the buffer solution to prepare reagent I;

[0049] 2), heating and boiling chloroauric acid and trisodium citrate according to the mass ratio of 1:1 to obtain a colloidal gold solution; adding the MYO antibody to the colloidal gold solution, and adding stabilizers, buffers and preservatives to obtain a reagent II; Wherein, when configuring chloroauric acid and trisodium citrate solution, use 0.2 μm filter membrane to filter;

[0050] 3) Reagent I and Reagent II are used together at a volume ratio of 5:1 to obtain a myoglobin detection reagent.

Embodiment 1

[0053] Preparation of colloidal gold particles (all the following operations need to be completed in a high cleanliness and dust-free space)

[0054] 1) Preparatory work: 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 goldware used, rinsed with distilled water, and set aside;

[0055] 2) Prepare a solution of 1% (w / v) chloroauric acid and 1% (w / v) sodium citrate, and filter it with a 0.2 μm filter membrane;

[0056] 3) Put a magnetic stirrer in a cleaned 1000ml round bottom flask, and add 1000ml ultrapure water;

[0057] 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 gr...

Embodiment 2

[0061] Preparation of reagent one

[0062] 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 myohemoglobin detection reagent which is formed by matching 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 20-50mmol / L of buffer solution, 2 percent (w / v) of a coagulant, 2 percent (w / v) of BSA and 0.1-0.5 percent (w / v) of a preservative; the reagent II comprises 0.1-0.5 percent (w / v) of colloidal gold particles labeled with anti-human MYO antibodies, 2-5 percent (w / v) of a stabilizer, 20-50mmol / L of buffer solution and 0.1-0.5 percent (w / v) of a preservative; and the diameters of the colloidal gold particles labeled with anti-human MYO antibodies are 60-90nm. The myohemoglobin detection reagent disclosed by the invention has the advantages of high analysis sensitivity, wide linear detection range, convenience in use, rapidness in detection and the like.

Description

【Technical field】 [0001] The invention relates to a myoglobin detection reagent and a preparation method thereof, in particular to a homogeneous sol particle type MYO immunoassay reagent for detecting MYO in serum or plasma and a preparation method thereof, belonging to the field of in vitro diagnostic reagents for medical devices. 【Background technique】 [0002] Myoglobin (MYO) is an oxygen-binding heme protein that is mainly distributed in cardiac and skeletal muscle tissues. It was elucidated by Kendrew with X-ray diffraction in 1960, which is the first tertiary structure of protein described in the world. In acute myocardial injury, MYO is first released into the blood. About 2 to 3 hours after the onset of symptoms, MYO in the blood can exceed the normal upper limit, reach the peak at 9 to 12 hours, and return to normal after 24 to 36 hours. Serial sampling is recommended for patients with suspected ACS because of the delay between the onset of symptoms and the release...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/72G01N33/577
CPCG01N33/577G01N33/721
Inventor 王会中
Owner 中国人民解放军第三0五医院
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