Folic acid detection kit and detection method

A detection kit and folic acid technology, applied in the field of medical analysis, can solve the problems of reducing the precision, reliability, insufficient reaction, magnetic bead agglutination and other problems of the detection results, and achieve the detection method is simple and easy to operate, no radioactive pollution stable effect

Active Publication Date: 2020-06-19
AUTOBIO DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this severe denaturation method will affect subsequent reactions, such as protein precipitation due to a sudden drop in pH, resulting in aggregation of magnetic beads; at the same time, excessive reducing agent damages horseradish peroxidase, resulting in insufficient reaction, thereby reducing detection The precision and reliability of the results, etc.

Method used

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  • Folic acid detection kit and detection method
  • Folic acid detection kit and detection method
  • Folic acid detection kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Preparation of the kit of the present invention

[0043] 1. Preparation of sample treatment solution 1 Add 24g of analytical pure NaOH to 1L of deionized water, stir evenly, and store at 2-8°C.

[0044] 2. Preparation of sample processing solution 2

[0045] Add 0.5% DTT to 0.05M, pH5.5 acetate buffer, stir evenly, and store at 2-8°C.

[0046] 3. Preparation of sample treatment solution 3

[0047] Add 1% iodoacetamide to 0.2M, pH5.0 MES buffer, stir evenly, and store at 2-8°C.

[0048] 4. Preparation of Magnetic Microparticle Suspension Coated with Folic Acid Derivatives

[0049] Fully mix the stock solution of carboxylated magnetic particles with a particle size of 1 μm, take out an appropriate amount for magnetic separation, remove the supernatant, add 0.05M, PH5.5 acetic acid buffer for re-suspending and washing, repeat twice, add the cross-linking agent EDC 10mg was used to activate the carboxyl groups on the surface of the magnetic particles, 1 μg of ...

Embodiment 2

[0054] Embodiment 2 The detection method of the kit of the present invention

[0055] (1) Collect 5 mL of whole blood using a non-added ordinary tube / separating rubber tube / promoting tube, let stand at 37°C for 30 minutes, and then centrifuge at 4000 rpm for 10 minutes, and take the centrifuged serum as the sample to be tested. (Note: The samples should not be stored at room temperature for more than 8 hours after collection. If the samples are not tested within 8 hours, they should be placed in a refrigerator at 2-8°C. If they need to be stored or transported for more than 48 hours, they should be frozen at -20°C. Below, avoid repeated freezing and thawing. Before use, return to room temperature and shake gently to mix).

[0056] (2) Treat the sample to be tested with sample treatment solution 1, sample treatment solution 2 and sample treatment solution 3 in turn, take 150 μl of the treated sample to be tested (or folic acid calibrator) and add 20 μL of magnetic particle susp...

Embodiment 3

[0057] Example 3 Performance testing of the kit of the present invention

[0058] Calibration curve: take the log value of the calibrator concentration value as the X axis, and take the log value of the calibrator luminous intensity value as the Y axis, calculate by four-parameter fitting method, and establish a calibration curve. The fitted curve is as follows figure 1 shown.

[0059] 1. Precision

[0060] Use the same batch number Example 1 kit to detect high and low indoor quality control products (Q1, Q2) respectively, and it is required to repeat the measurement for 4 days, at least 5 times a day, and fit the standard curve according to the four parameters (lin-log). Calculate the measurement results, then calculate the mean M and standard deviation SD of at least 20 measurement results, and obtain the coefficient of variation CV according to the formula CV=SD / M*100%. The test results are shown in Table 1 below.

[0061] Table 1

[0062]

[0063] It can be seen fro...

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Abstract

The invention relates to the technical field of medical analysis, in particular to a folic acid detection kit and a folic acid detection method. The kit comprises a magnetic particle suspension coatedwith a folic acid derivative, enzyme-labeled folic acid binding protein, a sample treatment solution and a chemiluminescent substrate, and the sample treatment solution comprises a sample treating fluid 1, a sample treating fluid 2 and a sample treating fluid 3. According to the kit, a specific sample treatment solution is adopted to treat a sample to be detected, folic acid and protein can be mildly separated, protein precipitation is not generated in the detection process, horse radish peroxidase is not damaged, a detection result is high in precision, the correlation with a Rogowski kit ishigh, and the detection result is accurate and reliable; and the detection speed is high, the result can be obtained within 40 minutes, and the kit is suitable for clinical detection of folic acid content.

Description

technical field [0001] The invention relates to the technical field of medical analysis, in particular to a folic acid detection kit and a detection method. Background technique [0002] Folic acid is a general term for a group of compounds with similar chemical structures and similar biochemical characteristics, consisting of pteridine, p-aminobenzoic acid and one or more glutamic acids. In its natural state, folic acid can exist in various forms, all of which can express its biological function in the form of isozymes, that is, the transport of single carbon units, which is essential for normal cell growth and DNA synthesis. [0003] The folic acid ingested by humans is folic acid polyglutamic acid, which needs to be hydrolyzed into folic acid polyglutamic acid at the brush border of the jejunal mucosa before it can enter the systemic circulation. Under the action of methylenetetrahydrofolate reductase, 5,10-methylenetetrahydrofolate is converted into 5-methyltetrahydrofo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76
CPCG01N21/76Y02A50/30
Inventor 谢茜尹伊郜志颖
Owner AUTOBIO DIAGNOSTICS CO LTD
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