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Preparation method of magnetic immunochromatographic test strip for quantitatively detecting alpha-fetoprotein in blood

A magnetic immunochromatography, alpha-fetoprotein technology, applied in the field of medical testing, can solve the problems of narrow linear range, operator harm, difficult to use in a large area, etc., and achieve the effects of simplified preparation process and easy operation.

Active Publication Date: 2012-10-24
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The current diagnostic techniques for AFP in blood mainly include: radioimmunoassay (RIA), enzyme-linked immunoassay (ELISA), chemiluminescence (CLIA), time-resolved fluorescence, colloidal gold immunochromatography and other methods, but these methods have their own Features and disadvantages: radioimmunoassay is a commonly used method in clinical practice. Its advantages are relatively accurate results and wide linear range. The disadvantages are cumbersome and time-consuming operations, and radioactive markers will cause harm to the operator and cause environmental pollution. It has been gradually replaced by other methods
ELISA is similar to chemiluminescence and time-resolved fluorescence method in principle, but there is a difference in sensitivity due to the difference in the final detection system. At present, automation, large-scale, and quantitative detection have been realized, but there are large differences in results between methods, and the linear range is relatively small. At the same time, automated testing is currently monopolized by large foreign manufacturers, and the equipment is expensive, and it is not suitable for single-person and small-batch testing, which greatly limits their application in primary hospitals and clinics.
In recent years, colloidal gold immunochromatography has appeared to detect AFP, which is fast, convenient, and single-serve operation, but the disadvantage is that it can only achieve semi-quantitative, can only indicate a general range, and cannot achieve accurate quantification, and abnormal samples need to use other methods Double-check testing will virtually increase the cost of medical treatment for the testees, and it is difficult to promote in the market, and it is difficult to use it on a large scale

Method used

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  • Preparation method of magnetic immunochromatographic test strip for quantitatively detecting alpha-fetoprotein in blood
  • Preparation method of magnetic immunochromatographic test strip for quantitatively detecting alpha-fetoprotein in blood
  • Preparation method of magnetic immunochromatographic test strip for quantitatively detecting alpha-fetoprotein in blood

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Embodiment 1

[0035] Preparation method of magnetic immunochromatography test strip and test box for quantitative detection of alpha-fetoprotein in blood

[0036] The preparation method of test strip and test paper box of the present embodiment comprises the following steps:

[0037] A. Antibody preparation: commercial AFP paired antibody (Hytest company, cat: 4F16) was selected, dialyzed against 20 mM PBS at pH 7.2 (pH7.0-7.6 is applicable) overnight at 4°C for later use.

[0038] B. Preparation of coating film:

[0039] Preparation of coating buffer: 0.02M phosphate buffer (PB) with pH 7.2 was used as coating buffer, sterilized by 0.22 μm microporous membrane filter and stored at 4°C for later use, valid for two weeks.

[0040] Preparation of blocking solution: 0.02M phosphate buffered saline (PBS) containing 0.5% BSA, pH 7.2 (pH 7.0-7.6 is applicable), 0.22μm microporous membrane filter sterilization, and store at 4°C for later use , valid for one week.

[0041] Preparation of coating...

Embodiment 2

[0070] Except in the step of the preparation of streptavidinized magnetic particles: streptavidin makes the ratio of streptavidin and magnetic particles 1:2, other steps are the same as in Example 1,

Embodiment 3

[0072] Except for the step of preparing streptavidinized magnetic particles: streptavidin makes the ratio of streptavidin to magnetic particles 1:10, other steps are the same as in Example 1.

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Abstract

The invention relates to a magnetic immunochromatographic test strip for quantitatively detecting alpha-fetoprotein in blood and a preparation method thereof. The test strip is assembled by pasting a coated film, magnetic particles combined with an AFP antibody, a sample pad and a water-absorbing pad which are mutually staggered by 2 millimeters in turn on a bottom plate and then covering an upperlayer with a transparent plastic sealing film, wherein the coated film is precoated with an AFP-antibody detection line and a quality control line. As the test strip introduces a magnetic immunochromatography technique and a biotin-avidin system into the quantitative detection of AFP in blood, the test strip has the advantages of greatly increasing detection sensitivity, providing accurate quantitative results, reducing the labor intensity of operators and ensuring that clinicians can quickly diagnose patients.

Description

technical field [0001] The invention belongs to the field of medical testing, in particular to a magnetic immunochromatographic test strip for quantitatively detecting alpha-fetoprotein in blood and a preparation method thereof. Background technique [0002] Alpha-fetoprotein (Alpha-fetoprotein, AFP) is produced by human liver and yolk sac (fetal) and is composed of 18 kinds of amino acids, carbohydrates account for about 4%, and the relative molecular weight is 65000-72000. glycoprotein. The content of alpha-fetoprotein varies greatly in different stages of the human life cycle. It is very high in fetal blood. It is synthesized by liver and yolk sac in the early stage of fetal development, and a small amount is synthesized by fetal vitellin cells and fetal gastrointestinal tract. - Synthesis begins at 10 weeks, reaches peak at 12-15 weeks, and a small amount passes through the placenta to maternal blood to increase AFP. After the fetus is born, alpha-fetoprotein decreases...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/558G01N33/577
Inventor 姚洪涛应希堂宋胜利胡国茂郑金来唐宝军于尚永
Owner CHEMCLIN DIAGNOSTICS CO LTD
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