Chemiluminescence kit for detecting vitamin B12

A chemiluminescent reagent, B12 technology, applied in the direction of biological testing, measuring devices, scientific instruments, etc., can solve the problems of low accuracy and long time consumption, and achieve the effect of saving time, simple operation and great clinical application value

Inactive Publication Date: 2019-11-12
NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a chemiluminescence kit for detecting vitamin B12, which can quantitatively detect vitamin B12 in human serum and plasma, and solves the problems of long time-consuming and low accuracy

Method used

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  • Chemiluminescence kit for detecting vitamin B12
  • Chemiluminescence kit for detecting vitamin B12
  • Chemiluminescence kit for detecting vitamin B12

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Chemiluminescence kit for detection of vitamin B12, including streptavidin magnetic bead suspension, vitamin B12 standard, biotin-labeled vitamin B12 monoclonal antibody, alkaline phosphatase-labeled vitamin B12 monoclonal antibody, chemiluminescence substrate , standard diluent, antibody diluent, washing solution, 96-well microtiter plate.

[0030] The streptavidin magnetic bead suspension is 10ml, and the concentration is 0.1mg / ml. The vitamin B12 standard product is a white freeze-dried powder with a concentration of 1 μg / ml. The volume of the biotin-labeled vitamin B12 monoclonal antibody is 0.2ml, and the concentration is 1mg / ml. The vitamin B12 monoclonal antibody labeled with alkaline phosphatase is 0.2ml, and the concentration is 1mg / ml. The chemiluminescence substrate (Lumi-phos530) is 100ml. The standard substance diluent contains 1% bovine serum albumin, 0.02% Procline-300 phosphate buffer (0.01M, pH7.4), and has a capacity of 10ml. The antibody diluent c...

Embodiment 2

[0033] Use the kit in Example 1 to detect the vitamin B12 standard substance, the method is as follows:

[0034] (1) The biotin-labeled vitamin B12 monoclonal antibody is the primary antibody, diluted 1 / 2000 times with antibody diluent.

[0035] (2) Alkaline phosphatase-labeled vitamin B12 monoclonal antibody is the secondary antibody, and the antibody diluent is diluted 1 / 2000 times.

[0036] (3) Dilute the vitamin B12 calibrator with standard diluent, and the concentrations are 10, 5, 2, 1, 0.5, 0.2, 0.1 ng / ml in turn.

[0037] (4) 50 μl of primary antibody, 50 μl of secondary antibody and 100 μl of vitamin B12 calibrator were added to the 96-well ELISA plate.

[0038] (5) After shaking and mixing, incubate in a 37°C incubator for 60 minutes.

[0039] (6) Add 100 ul of streptavidin magnetic bead suspension to each well at a concentration of 0.1 mg / ml.

[0040] (7) After shaking and mixing, incubate for 30 minutes in a 37°C incubator.

[0041] (8) Take out the ELISA plate...

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Abstract

The invention discloses a chemiluminescence kit for detecting vitamin B12. The chemiluminescence kit for detecting vitamin B12 comprises a streptavidin magnetic bead suspension, a vitamin B12 standardsubstance, a biotin-labeled vitamin B12 monoclonal antibody, an alkaline phosphatase-labeled vitamin B12 monoclonal antibody, a chemiluminescent substrate, a standard substance diluent, an antibody diluent, a washing solution, and a 96-well ELISA plate. The chemiluminescence kit for detecting vitamin B12 utilizes the technical characteristics of magnetic beads, and combines the biotin-avidin enzyme-linked immunosorbent amplification and a alkaline phosphatase-labeled enzymatic chemiluminescence system to detect the vitamin B12 level in serum, which has very high sensitivity; the chemiluminescence kit for detecting vitamin B12 has more technical and price advantages compared with the prior art; and the chemiluminescence kit for detecting vitamin B12 has the advantages of simple operation,time saving, easy realization of fully automatic operation, no need of expensive special instruments, relatively low requirements on the technical level of the operator and laboratory conditions, which is suitable for promotion and use in the primary hospital, and has great clinical application value.

Description

technical field [0001] The invention relates to the field of in vitro diagnosis, in particular to a chemiluminescent kit for detecting vitamin B12. Background technique [0002] Vitamins are a class of low-molecular organic compounds necessary for various organisms to maintain normal physiological functions. Most of them are the main coenzymes in the process of biochemical metabolism. They participate in, regulate, and control many synthesis and catabolism in the body. When organisms lack vitamins , It will cause substance metabolism disorders and cause various diseases. Vitamin B12, also known as cobalamin, is a light red needle-like crystal, easily soluble in water and ethanol, most stable under weak acid conditions with a pH value of 4.5 to 5.0, decomposed in strong acids (pH<2) or alkaline solutions, and heated It can be damaged to a certain extent, but the loss of short-term high-temperature disinfection is small, and it is easily destroyed by strong light or ultrav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/82G01N33/543
CPCG01N33/82G01N33/543
Inventor 周义正唐静陈星星黄丹娣董文霞
Owner NINGBO AUCHEER BIOTECHNOLOGY CO LTD
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