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A kind of neopterin nano magnetic particle chemiluminescent immunoquantitative detection kit and its preparation method

A technology of chemiluminescent immunity and nano-magnetic particles, which is applied in the field of immunoanalysis medicine, can solve the problems of poor repeatability of measured values, narrow detection range, and low sensitivity of enzyme-linked immunoassay, and achieve good performance, good stability, and low cost. Effect

Active Publication Date: 2015-09-02
BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The problem to be solved by the present invention is to provide a chemiluminescence immunoassay kit for neopterin and its preparation method, 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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  • A kind of neopterin nano magnetic particle chemiluminescent immunoquantitative detection kit and its preparation method
  • A kind of neopterin nano magnetic particle chemiluminescent immunoquantitative detection kit and its preparation method

Examples

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Effect test

Embodiment 1

[0042] Example 1: Preparation of Neopterin Nano Magnetic Particle Chemiluminescent Immunoquantitative Assay Kit

[0043] (1) Preparation of neopterin calibrator:

[0044] Dilute the anti-neopterin anti-pure product (purchased from US BIO company) calibrator diluent to the working concentration, respectively 0, 1, 5, 20, 80, 250nmoL / L;

[0045]Described calibrator diluent is the phosphate buffer saline containing 1%BSA;

[0046] (2) Preparation of neopterin quality control substances:

[0047] Dilute pure neopterin to 3nmoL / L and 200nmoL / L with calibrator diluent, 3nmoL / L is used as a low-value quality control product, and 200nmoL / L is used as a high-value quality control product;

[0048] (3) Preparation of magnetic nanoparticle-streptavidin suspension:

[0049] A. Preparation of ferroferric oxide nano-magnetic particles

[0050] Precipitation method was used to prepare ferroferric oxide nano-magnetic particles. The specific preparation method was as follows: 1) FeCl 3 ·6...

Embodiment 2

[0080] Embodiment 2: the inspection of kit of the present invention

[0081] (1) Physical inspection: liquid components should be clear without sediment or floc; other components should have no package damage.

[0082] (2) 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 curve within the concentration range of 0-2500nmoL / L is not less than 0.9900.

[0083] (3) Analytical sensitivity: The analytical sensitivity of the kit is not higher than 0.5nmoL / L.

[0084] (4) Precision: 10 wells were measured in parallel for high-value and low-value quality control products, and the average concentration of the measurement results was calculated ( ) and standard deviation (SD), within-assay imprecision Use 3 batches of products to carry out 3 tests, calculate the average concentration of assay result ( ) and standard deviation (SD), inter-assay imprecision The results...

Embodiment 3

[0087] Embodiment 3: the using method of kit of the present invention

[0088] (1) Equilibrate the test kit at room temperature (18-25°C) for 30 minutes.

[0089] (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.

[0090] (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.

[0091] (4) Number the reaction tubes, add 20uL calibrator or serum sample, 50uL magnetic particle-streptavidin suspension, 50uL biotin-neopterin antibody conjugate, 100uL neopterin antibody enzyme conjugate to the test tube in sequence 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 t...

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Abstract

The present invention discloses a neopterin (NPT) nano-magnetic particle chemiluminescent immunity quantitative detection kit. The kit comprises: a neopterin calibrator; a nano-magnetic particle suspension solution coupled with streptavidin; a biotin-labeled neopterin antibody; a neopterin antibody enzyme conjugate, with the used enzyme being horseradish peroxidase, which has purity RZ of greater than or equal to 3.0 and activity of greater than or equal to 250U / mL; a neopterin quality control product; a chemiluminescent liquid A and a chemiluminescent liquid B; a 20-time concentrated lotion; and reaction tubes. In addition, the invention also discloses a preparation method of the kit. Compared with the existing kits, the kit provided in the invention has the advantages of high sensitivity, wide measurable concentration range, long effective period of reagents, low reagent cost, simple operation, and high detection automation degree.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine. Specifically, the invention provides a neopterin (NPT) nano-magnetic particle chemiluminescent immunoquantitative detection kit and a preparation method thereof. Background technique [0002] Neopterin (neopterin, NPT) is the most important pteridine substance in human body fluids, and its chemical name is 2-amino-4-hydroxy-6-(D-erythrose Propyl)pterin, which is a low-molecular compound derived from guanosine triphosphate (GTP) metabolism and an intermediate metabolite in the biosynthesis of tetrahydrobiopterin. It is a pteridine compound produced by IFN-γ stimulation of macrophages. Neopterin is a sensitive marker of cell-mediated immune activation, and its concentration increases in diseases related to cell-mediated immune activation. The detection of neopterin is mainly used in tumors, trauma, immune system diseases, endocrine system diseases, and digestive system diseases , circulator...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N21/76G01N33/531
Inventor 刘萍栾大伟李克锦
Owner BIOSCIENCE (TIANJIN) DIAGNOSTIC TECH CO LTD