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Magnetic bead immunochromatographic test strip for quickly detecting tropomyosin and application thereof

An immunomagnetic bead chromatography, tropomyosin technology, applied in the detection field, can solve the problems of complicated processing, increased operation time, unfavorable rapid detection and the like

Active Publication Date: 2018-02-23
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Zhongsheng Beikong Biotechnology Co., Ltd. invented a colloidal gold test strip for detecting aquatic allergens, which is characterized by a carrier plate and nitrocellulose membrane (patent number CN201410231103.9), but its sample pretreatment Complexity is not conducive to rapid on-site detection
Shanghai Ocean University invented a magnetic immunochromatographic method for rapid detection of tropomyosin and the preparation of test strips (patent number CN102043055.A). The quantitative detection of tropomyosin with a magnetic signal meter can reach 0.15 μg / mL, but Adding immunomagnetic beads to the sample increases the operating time during the detection, and the detection is performed with the help of an instrument, and the pretreatment method is complicated and time-consuming

Method used

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  • Magnetic bead immunochromatographic test strip for quickly detecting tropomyosin and application thereof
  • Magnetic bead immunochromatographic test strip for quickly detecting tropomyosin and application thereof
  • Magnetic bead immunochromatographic test strip for quickly detecting tropomyosin and application thereof

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Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1 prepares test strip

[0027] 1. Prepare the binding pad: the binding pad is a binding pad labeled with magnetic beads modified by anti-Tm specific murine monoclonal antibody; dissolve 60 μl of magnetic beads labeled with the antibody in 1680 μl of the sample pad treatment solution and the binding pad treatment solution In the mixed solution (sample pad treatment solution: conjugation pad treatment solution at a ratio of 1:2), the mixture was evenly added dropwise on the conjugation pad at a rate of 70 μl / cm. When dropping, it should be suspended in the air and added dropwise evenly. Dry for two hours after the dropwise addition.

[0028] 2. Preparation of test strips: the test strips are pasted on the bottom plate in turn by absorbent paper, nitrocellulose membrane, binding pads, and sample pads from top to bottom after the binding pads are dried; the nitrocellulose membranes are pre-coated Nitrocellulose membrane with test line for Tm detection antigen a...

Embodiment 2

[0038] (1) Prepare acetone and shrimp meat according to 1g: 5ml, extract with acetone to colorless shrimp meat, add Tris-HCl buffer solution (containing 0.5mol / L NaCl, pH 9.2), use an electric homogenizer to homogenize for 30 s, and the resulting mixture is subjected to ultrasonic treatment for 10 min, then heated for 5 min, cooled rapidly, and the resulting solution is centrifuged for 10 min in a mini centrifuge, and the supernatant is used for detection;

[0039] The supernatant was subjected to SDS-PAGE and Western-blooting detection to determine the purity and content of the extracted tropomyosin, see appendix image 3 shown. It can be seen that the samples processed by this method have fewer bands of miscellaneous proteins, and the obtained tropomyosin has a high content.

[0040] (2) The obtained supernatant solution is the sample to be tested. Add 120 μL of the sample to the sample pad of the test strip, and observe the result after 8 minutes of chromatography. The res...

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Abstract

The invention belongs to the field of detection, particularly relates to tropomyosin in crustacean aquatic products such as shrimps, crabs and other molluscs, and discloses a sample pretreating methodsuitable for quickly detecting tropomyosin in the crustacean aquatic products on site and a detection reagent box thereof. A test strip comprises a bottom plate, a sample pad, a binding pad, a nitrocellulose membrane and a water absorbing pad; the sample pad, the binding pad, the nitrocellulose membrane and the water absorbing pad are sequentially pasted to the upper surface of the bottom plate in a staggered mode from one end to the other end; the binding pad is a binding pad marked by magnetic beads modified by anti-tropomyosin specificity rat monoclonal antibodies; the nitrocellulose membrane is a nitrocellulose membrane precoated with a detection line for Tm detection antigen and a control line for goat anti-mouse IgG. By means of the sample pretreating method suitable for quickly detecting tropomyosin in the crustacean aquatic products on site and the quick detection reagent box thereof, the content of tropomyosin in a detected sample can reach 5 microgram / mL within a visual range.

Description

technical field [0001] The invention belongs to the detection field, and in particular relates to tropomyosin in aquatic products such as crustacean aquatic products, shrimps, crabs and molluscs, and is applicable to a sample pretreatment method and a detection kit for rapid on-site detection of tropomyosin in crustacean aquatic products. Background technique [0002] Shrimp, crab and lobster in aquatic products are the main allergens of crustaceans, and tropomyosin is the most important allergen. The allergic symptoms caused by them are becoming more and more complicated and serious, and traditional cooking methods cannot degrade tropomyosin , so it often causes food poisoning incidents; the current detection technology uses mass spectrometry technology to analyze the amino acid sequence of the whole protein or the peptide after enzymatic hydrolysis, polymerase chain reaction (Polymerase Chain Reaction, PCR) technology and analysis of DNA fragments encoding allergens Allerg...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/577G01N33/558G01N33/543G01N1/28
CPCG01N1/28G01N33/54326G01N33/558G01N33/577G01N33/68G01N2333/4712
Inventor 卢瑛林昕
Owner SHANGHAI OCEAN UNIV
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