Magnetic immune chromatography method capable of synchronously detecting Tm and Pa allergens

A technology of magnetic immunochromatography and simultaneous detection, which is applied in clinical detection of allergens, food inspection, and detection of food allergens. The effect of strong stability

Inactive Publication Date: 2011-08-17
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] Purpose of the present invention: aiming at the insufficiency and defect that the prior art cannot simultaneously detect allergen Tm and Pa, provide a kind of immunochromatographic method using immunomagnetic beads as markers, which can capture Tm or Pa allergens alone, and can also Capture Tm and Pa allergens at the same time, so as to realize the purpose of simultaneous detection of Tm and Pa, the main allergens of aquatic products

Method used

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  • Magnetic immune chromatography method capable of synchronously detecting Tm and Pa allergens
  • Magnetic immune chromatography method capable of synchronously detecting Tm and Pa allergens
  • Magnetic immune chromatography method capable of synchronously detecting Tm and Pa allergens

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preparation example Construction

[0069] The preparation method of the magnetic immunochromatography test paper strip which can detect Tm and Pa allergen simultaneously comprises the following steps:

[0070] C. Preparation and purification of antigen: Tm allergen uses shrimp as raw material, Pa allergen uses crucian carp as raw material

[0071] Raw materials were prepared and purified by methods designed by the laboratory.

[0072]D. Preparation and purification of antibodies: First, multiply the specific antibody cell lines of Tm and Pa allergens

[0073] cultured, and then injected into BALB / C mice to prepare ascites; the prepared ascites was concentrated with 50% (w / v) ammonium sulfate and then used commercial protein-G affinity chromatography column, according to the attached product instructions Purify.

[0074] C. Preparation of immunomagnetic beads: select magnetic beads with a diameter of 50-200 nm, and use carbodimethylamide (EDC) and succinimide (NHS) to covalently cross-link anti-Tm monoclonal a...

Embodiment 1

[0091] Embodiment 1: Magnetic immunochromatography detects the main allergen Tm of crustaceans

[0092] (1) Preparation and purification of main crustacean allergen Tm and fish main allergen Pa: take shrimp acetone

[0093] 2g powder, add 30mL 20mmol / L Tris-HCl (pH 7.5) solution containing 1 mol / L KCl to extract overnight, after centrifugation, take the supernatant and put it in a water bath at 100 ℃ for 10min, then centrifuge to recover the supernatant. The resulting supernatant was treated by PI 4.5 isoelectric point precipitation, and the precipitate was dissolved in 20mmol / L Tris-HCl (pH 7.5), adjusted to pH 7.5 with 1 mol / L NaOH, and subjected to 40% ammonium sulfate salting out, and recovered after centrifugation The supernatant was subjected to the isoelectric point operation and salted out with 60% ammonium sulfate. After centrifugation, the precipitate was dissolved in PBS and stored at -20°C for later use.

[0094] The purification method of the main fish allergen P...

Embodiment 2

[0101] Embodiment 2: Magnetic immunochromatography detects fish main allergen Pa

[0102] Except that in the sample detection step, the detection sample is the purified main fish allergen Pa, other steps are the same as in Example 1.

[0103]

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Abstract

The invention discloses a magnetic immune chromatography method capable of synchronously detecting Tm and Pa allergens. The method is characterized by comprising the following steps of: sequentially sticking a sample pad, a combination pad combining anti-Tm and anti-Pa immune magnetic beads, a chromatography film and a water absorption pad to a bottom plate crossly at intervals of about 2 millimeters, and then covering a transparent plastic sealing film on the upper layer to construct a magnetic immune chromatography test strip capable of synchronously detecting the Tm and Pa allergens. A Pa antigen detection line T1, a Tm antigen detection line T2 and a goat anti-mouse IgG quality control line C are pre-coated on the chromatography film, the immune magnetic beads can be captured by chromatography, and quick qualitative detection of the Tm and Pa allergens is realized according to 1 to 3 macroscopic color development strips formed after the chromatography of a sample; or quantitative detection of single allergen or synchronous quantitative detection of two allergens is realized by detecting the constructed magnetic immune test strips through a magnetic analyzer according to the magnetic signal detection values of the detection lines and the control line formed after the chromatography of the sample.

Description

technical field [0001] The invention relates to a magnetic immunological detection technology for allergens, in particular to a magnetic immunochromatographic method capable of synchronous detection of Tm and Pa allergens, which belongs to the field of food inspection, relates to the technical field of detection of food allergens, and also relates to allergens The original field of clinical testing. Background technique [0002] Food allergy is a worldwide public health problem. Surveys show that about 4% of the world's population is allergic to food. In order to protect the health of susceptible people, some countries and regions have strictly regulated the labeling of food allergens and included them in the scope of legislation. Therefore, the detection of food allergens is becoming more and more important. A food allergy is an abnormal reaction of the body's immune system to food. A food allergen is a substance in food that triggers or provokes an allergic reaction. It ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/532G01N33/543
Inventor 卢瑛石良王锡昌
Owner SHANGHAI OCEAN UNIV
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