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Kit for chemiluminescence immunity quantitative detection of MYO (myohaemoglobinnano) nano magnetic particle and preparation method of kit

A technology of chemiluminescent immunity and nano-magnetic particles, which is applied in the field of immunoanalysis medicine, can solve the problems of narrow detection range, inaccurate quantification of colloidal gold test strips, and poor sensitivity of enzyme-linked immunoassay, achieving good stability and low cost , good specific effect

Active Publication Date: 2013-09-04
BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The problem to be solved by the present invention is to provide a chemiluminescence immunoassay quantitative detection kit for myoglobin and a preparation method thereof, which avoids defects such as inaccurate quantification of colloidal gold test strips, poor sensitivity of ELISA, and narrow detection range.

Method used

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  • Kit for chemiluminescence immunity quantitative detection of MYO (myohaemoglobinnano) nano magnetic particle and preparation method of kit
  • Kit for chemiluminescence immunity quantitative detection of MYO (myohaemoglobinnano) nano magnetic particle and preparation method of kit

Examples

Experimental program
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Effect test

Embodiment 1

[0058] Example 1: Preparation of Myoglobin Nano Magnetic Particle Chemiluminescent Immunoquantitative Assay Kit

[0059] (1) Preparation of myoglobin calibrator:

[0060] The myoglobin antigen (purchased from Fitzgerald Company) was prepared with goat serum (Zhengzhou Yikang Bioengineering Co., Ltd.) to make a concentrated stock solution of calibrator, and the enterprise standard was used for calibration, and the concentrated stock solution was diluted to the working concentration with goat serum , respectively 0, 5, 25, 100, 250, 1000ng / mL;

[0061] (2) Preparation of myoglobin control product:

[0062] Dilute the above concentrated stock solution to 10ng / mL and 500ng / mL with goat serum, use 10ng / mL as a low-value quality control, and 500ng / mL as a high-value quality control;

[0063] (3) Preparation of magnetic nanoparticle-streptavidin suspension:

[0064] A. Preparation of ferroferric oxide nano-magnetic particles

[0065] Precipitation method was used to prepare ferro...

Embodiment 2

[0081] Embodiment 2: the inspection of kit of the present invention

[0082] (1) Physical inspection: liquid components should be clear without sediment or floc; other components should have no package damage.

[0083] (2) Accuracy: The kit calibrator and the national standard series are analyzed and measured at the same time, and the double-logarithmic mathematical model is used to fit the two dose-response curves. It is required that the two dose-response curves do not deviate significantly from parallel (t test, |t|<2.447) ; Take the myoglobin enterprise standard as the reference substance, and use the double-logarithmic mathematical model to fit, and the average value of the ratio between the measured value and the labeled value of the kit calibrator should be within the range of 0.90 to 1.10.

[0084] (3) Linearity of the dose-response curve: Fitting with a double-logarithmic mathematical model, the absolute value of the correlation coefficient r of the dose-response curv...

Embodiment 3

[0091] Embodiment 3: the using method of kit of the present invention

[0092] (1) Equilibrate the test kit at room temperature (18-25°C) for 30 minutes.

[0093] (2) Preparation of lotion: dilute the concentrated lotion with distilled water at 1:20 (1mL of lotion plus 19mL of distilled water). If the concentrated lotion has crystals, the concentrated lotion can be placed at room temperature or 37°C, and then diluted after the crystals dissolve.

[0094] (3) Preparation of luminous liquid: Take an appropriate amount of luminous liquid A and luminous liquid B and mix them in equal volumes 5 minutes before use.

[0095] (4) Number the reaction tubes, add 25uL calibrator or serum sample, 50uL magnetic particle-streptavidin suspension, 50uL biotin-myoglobin antibody conjugate, 100uL myoglobin antibody enzyme conjugate to the test tube Shake and react at 37°C for 30 minutes, place the test tube rack on a magnetic separator for separation for 5 minutes, then pour out the supernata...

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Abstract

The invention discloses a kit for chemiluminescence immunity quantitative detection of an MYO nano magnetic particle. The kit comprises an MYO calibrator, a nano magnetic particle suspension liquid coupled with streptavidin, bioti-labeled MYO antibodies, MYO abzyme conjugate, an MYO quality control product, a chemiluminescence liquid A and a chemiluminescence liquid B, a 20-time concentrated washing liquor and a reaction tube, wherein for the MYO abzyme conjugate, used enzyme adopts horse radish peroxidase with purity RZ larger than or equal to 3.0 and activity larger than or equal to 250 U / mL. Besides, the invention further discloses a preparation method of the kit. Compared with the conventional kit, the kit provided by the invention has the advantages of high sensibility and test automation, wide measurable concentration range, long useful life of a reagent, simple operation and the like.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine. Specifically, the invention provides a myoglobin (MYO) nano magnetic particle chemiluminescent immunoquantitative detection kit and a preparation method thereof. Background technique [0002] Myoglobin (MYO) is a heme protein similar to hemoglobin. Exist in muscle cells, the main function is to store oxygen, and it is very abundant in the myocardium. The molecular weight is 16700, and it is composed of a polypeptide chain containing 153 amino acid residues and a heme prosthetic group, and is in a compact spherical shape. Most of the hydrophobic side chains on the amino acid residues in the polypeptide chain are inside the molecule, and the hydrophilic side chains are mostly on the surface of the molecule, so its water solubility is better. [0003] Due to the small molecular weight of MYO, it is released into the blood circulation early after muscle cell damage, increases in 1-2 hours, re...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/531G01N21/76
Inventor 刘萍栾大伟宋启超董婷婷李克锦
Owner BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD
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