Chemiluminiscence immuno-assay kit for 25-hydroxyl vitamin D and preparation method thereof

Abstract

The invention discloses a chemiluminiscence immuno-assay kit for 25-hydroxyl vitamin D and a preparation method thereof. The chemiluminiscence immuno-assay kit herein includes: carboxylated magnetic micro-particles that are coated by a 25-hydroxyl vitamin D monoclonal antibody and a chemiluminiscence marker that is marked by a vitamin D derivative. The chemiluminiscence immuno-assay kit, with a full-auto chemiluminiscence immuno-analysis instrument as a detection tool, completes the detection on the 25-hydroxyl vitamin D. A test proves that the kit reaches 0.2 ng / ml in detection sensitivity, which is improved by at least 10 times than that of a conventional detection method of the 25-hydroxyl vitamin D. The chemiluminiscence immuno-assay kit has high detection precision.

Description

technical field [0001] The invention relates to the field of in vitro detection, in particular to a 25-hydroxyvitamin D chemiluminescent immunoassay kit and a preparation method thereof. Background technique [0002] Vitamin D (vitamin D) is a sterol derivative with anti-rickets effect, also known as anti-rickets vitamin. Can be divided into vitamin D2 and vitamin D3. Vitamin D2 is mostly contained in plant foods. It is synthesized by plant ergosterol through sunlight. Vitamin D3 can be synthesized by human skin and fat tissue from 7-dehydrocholesterol through sunlight. Vitamin D2 is a fat-soluble vitamin. Vitamin D from food is absorbed through the small intestine together with fat, forms chylomicrons with the assistance of bile, enters the blood from lymphatic vessels, and is transported to the liver together with self-synthesized vitamin D3. In the liver, 25-hydroxyvitamin D is formed through the action of the monooxygenase system (25-hydroxylase) in the hepatocyte micr...

AI Technical Summary

Problems solved by technology

[0010] (1) Use 12×8 type, 6×8 type, 8×12 type or full-plate type 96-well special microwell plate as antigen coating equipment and reaction container, which can only be divided into 12 batches and 6 batches when used , 8 batches or the whole board can be used at one time, and independent and single-person testing cannot be carried out;

[0011] (2) There are many kinds of reagents used in quantitative determination, and each detection reagent must be contained in a reagent bottle, and each time a reagent is used, the suction nozzle needs to be replaced to fill the microwells of the microwell plate respectively , not only there are many types of reagent bottles, but also the operation of filling reagents is extremely cumbersome;

[0012] (3) There is a lack of corresponding labeling of the testing information, and the production batch number and expiration date information of the testing reagent can only be known or known by checking the label on the outer packaging box of the kit, and the known information is not controlled during the testing process, which has great potential large randomness;

[0013] (4) The detection reagent is in an open space during the detection process, which may easily cause cross-contamination between various reagents and affect the accuracy of the detection result;

[0014] (5) Manual operation is mostly used in the detection process, the addition of reagents or samples is not very precise, the operation process is extremely cumbersome and complicated, and operation errors are prone to occur, and the accuracy and precision of the detection results are poor;

[0015] (6) The quantity configuration and use of the complete set of reagents for the test items are the number of items × 48 / 96 persons. If 10 items need to be tested, the configuration and use of the reagents must be 10 × 48 / 96 persons. If Only one sample needs to detect 10 different items, and it also needs to configure reagents for 10×48 / 96 people, which has the disadvantage of not being economical and reasonable

[0018] Enzymatic chemiluminescence mainly includes horseradish peroxidase (HRP) and alkaline phosphatase, but both have certain limitations. The main disadvantage of horseradish peroxidase is: In the presence of biomolecules, it will also be blocked by H 2 o 2 Oxidation is self-luminescent, the background is relatively high, which affects the signal-to-noise ratio, the reaction kinetics is complex, there are many influencing factors, the result is not stable enough, and it is not easy to obtain a substrate with high sensitivity and long plateau period

The main disadvantages of alkaline phosphatase are: it takes a long time for the substrate to reach the plateau, and the cost of the substrate is high, resulting in high detection cost and heavy burden on patients

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