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Kit for chemiluminescence immunity quantitative detection of CK-MB (creatine kinase- isoenzyme) nano magnetic particle and preparation method of kit

A chemiluminescence immunoassay and nano-magnetic particle technology, which is applied in the field of immunoassay medicine, can solve the problems of narrow detection range, low sensitivity of enzyme-linked immunosorbent assay, poor repeatability of measured values, etc., and achieves good stability, good specificity and good performance. 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

[0009] The problem to be solved by the present invention is to provide a chemiluminescence immunoassay kit for creatine kinase isoenzymes and a preparation method thereof, which avoids defects such as poor repeatability of measured values, low sensitivity of enzyme-linked immunoassay, and narrow detection range.

Method used

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  • Kit for chemiluminescence immunity quantitative detection of CK-MB (creatine kinase- isoenzyme) nano magnetic particle and preparation method of kit
  • Kit for chemiluminescence immunity quantitative detection of CK-MB (creatine kinase- isoenzyme) nano magnetic particle and preparation method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1: Preparation of Creatine Kinase Isoenzyme Nano Magnetic Particle Chemiluminescent Immunoquantitative Assay Kit

[0057] (1) Preparation of creatine kinase isoenzyme calibrator:

[0058] Prepare creatine kinase isoenzyme antigen (purchased from Fitzgerald Company) with calibrator diluent (phosphate buffer solution containing 1% BSA) to prepare concentrated stock solution of calibrator, calibrate with enterprise standard, and dilute concentrated stock solution Dilute to the working concentration with calibrator diluent, respectively 0, 3, 15, 50, 200, 800ng / mL;

[0059] (2) Preparation of Creatine Kinase Isoenzyme Quality Control:

[0060] Dilute the above concentrated stock solution to 12ng / mL and 300ng / mL with calibrator diluent, 12ng / mL is the low quality control, 300ng / mL is the high quality control;

[0061] (3) Preparation of magnetic nanoparticle-streptavidin suspension:

[0062] A. Preparation of ferroferric oxide nano-magnetic particles

[0063] Pre...

Embodiment 2

[0079] Embodiment 2: the inspection of kit of the present invention

[0080] (1) Physical inspection: liquid components should be clear without sediment or floc; other components should have no package damage.

[0081] (2) Accuracy: The kit calibration product and the national standard product series are analyzed and determined at the same time, and the double-logarithmic mathematical model is used for fitting, requiring that the two dose-response curves do not deviate significantly from parallel (t test, |t|<2.447) ; Taking creatine kinase isoenzyme enterprise standard product as the reference substance, and using a double logarithmic mathematical model to fit, 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.

[0082](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-respon...

Embodiment 3

[0090] Embodiment 3: the using method of kit of the present invention

[0091] (1) Equilibrate the test kit at room temperature (18-25°C) for 30 minutes.

[0092] (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.

[0093] (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.

[0094] (4) Number the reaction tube, add 20uL calibrator or serum sample, 50uL magnetic particle-streptavidin suspension, 50uL biotin-creatine kinase isozyme antibody conjugate, and 100uL creatine kinase to the test tube in sequence Isozyme-antibody-enzyme conjugates, shake and react at 37°C for 30 minutes, place the test tube rack on a magnetic separator for separati...

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Abstract

The invention discloses a kit for chemiluminescence immunity quantitative detection of a CK-MB nano magnetic particle. The kit comprises a CK-MB calibrator; a nano magnetic particle suspension liquid coupled with streptavidin, bioti-labeled CK-MB antibodies, CK-MB abzyme conjugate, a CK-MB 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 CK-MB 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 validity 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 creatine kinase isoenzyme (CK-MB) nano magnetic particle chemiluminescent immunoquantitative detection kit and a preparation method thereof. Background technique [0002] Creatine phosphokinase (Creatine Kinase, CK) exists in large quantities in cardiac muscle, skeletal muscle and brain and other tissues. It is an important kinase that is directly related to intracellular energy operation, muscle contraction, and ATP regeneration. The natural CK molecule is a compact spherical structure, which is a dimer composed of two subunits (M-type subunit and B-type subunit). CK has three major isozymes, namely CK-MM, CK-MB, and CK-BB. Among them, CK-BB mainly exists in the brain, prostate, intestine, and lung; there are a large amount of CK-MM and a small amount of CK-MB in skeletal muscle; and although CK-MM is the main tissue in myocardial tissue, the proport...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/577G01N33/70
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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