Gold-labeled test strip/card for quickly detecting mercury ions

A gold-labeled test paper and mercury ion technology, which is applied to gold-labeled test paper strips/cards and the preparation method and application fields thereof, can solve the problems such as inability to meet the demand for cheap and fast on-site detection, poor selectivity, and inability to achieve

Inactive Publication Date: 2014-01-08
HENAN INST OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, most of the mature mercury detection technologies rely on large-scale analytical instruments, which cannot meet the needs of cheap and fast on-site detection.
However, commercial rapid detection instruments and mercury test strips have low sensitivity (the lower limit of detection is 0.1mg / L), which cannot reach the limit values ​​stipulated by the national standard (0.05mg / L for sewage, 0.001mg / L for drinking water), and Poor selectivity, unable to meet sensitive and accurate detection requirements

Method used

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  • Gold-labeled test strip/card for quickly detecting mercury ions
  • Gold-labeled test strip/card for quickly detecting mercury ions
  • Gold-labeled test strip/card for quickly detecting mercury ions

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Embodiment 1, mercury ion artificial immunization antigen preparation

[0035] Prepare artificial immune antigen mercury ion-chelating agent-carrier protein, with Hg 2+ - Take ITCBE-OVA as an example, weigh 20 mg of ovalbumin (OVA) and dissolve in 1 mL of HEPES buffer (10 mM / L) at pH 9.0 to form a 20 mg / mL carrier protein solution. Weigh 10 mg of 1-(4-isothiocyanatophenyl)-ethylenediaminetetraacetic acid (ITCBE) and dissolve it in 1 mL of dimethyl sulfoxide (DMSO) to form a metal chelating agent solution; add 0.5 mL of the metal chelating agent solution dropwise Add it to the OVA carrier protein solution, mix well, add 100ul tri-n-butylamine (1.5M), and react at room temperature on a shaker for 24h to prepare an OVA-ITCBE solution. Take 11.25mg mercury nitrate (Hg(NO 3 ) 2 ) was dissolved in 100ul distilled water to form a uniform Hg 2+ solution; the Hg 2+ Add the solution to the OVA-ITCBE solution, adjust the pH value to 7.4, incubate on a shaking table at room te...

Embodiment 2

[0038] Embodiment two, mercury ion artificial immune antigen identification

[0039] Determination of Hg by the biquinoline carboxylic acid method 2+ - Carrier protein OVA concentration in ITCBE-OVA. OVA was used as the standard protein, and the concentration gradients of 0 μg / mL, 10 μg / mL, 20 μg / mL, 40 μg / mL, 60 μg / mL, and 80 μg / mL were prepared, and the concentration detection standard curve was constructed by the biquinolinic acid method. The linear equation of the OVA concentration standard curve is: y=0.0035x+0.0065, R 2 =0.9981, where y is the absorbance of the sample at a wavelength of 562nm, and x is the protein concentration of the sample.

[0040] Determination of Hg by ICP-AES 2+ -ITCBE-OVA Hg 2+ concentration in . 100 μg / mL of Hg 2+ -Hg in ITCBE-OVA 2+ Dilute the standard stock solution with 2% nitric acid to a concentration gradient of 0 μg / mL, 0.1 μg / mL, 0.2 μg / mL, 0.4 μg / mL, 0.6 μg / mL, and 0.8 μg / mL, and the instrument software automatically draws a stand...

Embodiment 3

[0041] Embodiment 3, preparation of anti-mercury ion monoclonal antibody

[0042] with Hg 2+ -ITCBE-OVA immunized 5 female BALB / C mice for 6 weeks at a dose of 50μg / 0.2mL / mouse, subcutaneously injected on the back. Select spare mice for cell fusion, and detect Hg in antiserum by indirect ELISA 2+ Chelate pAb titer, blocking ELISA to detect Hg 2+ -ITCBE pAb vs Hg 2+ - IC of ITCBE 50 , select the highest titer, IC 50 Minimal mice, hyperimmunized for cell fusion. Use cell fusion technology to aseptically take spleen to prepare spleen cells, and fuse with NS0 myeloma cells under the action of 50% PEG (pH8.0); use indirect ELISA and blocking ELISA to screen positive hybridoma cell lines, and select strong positive Wells with high inhibition rate and strong cell growth were cloned by limiting dilution three times. Resuscitate the hybridoma cells after freezing for 15 days, 30 days and 60 days respectively, take the supernatant after culture, measure the antibody titer by indi...

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Abstract

The invention discloses a gold-labeled test strip / card for quickly detecting mercury ions. The gold-labeled test strip / card comprises a bottom support plate layer (1), a sample pad (2), a gold-labeled antibody binding pad (3), a cellulose membrane (4) and an absorbent pad (5). The gold-labeled test strip / card can quickly detect the residual contamination of the mercury ions in environments, soil, water, food, drugs and cosmetics.

Description

Technical field: [0001] The invention relates to a gold standard test strip / card for rapid detection of mercury ions, a preparation method and application thereof, and belongs to the technical fields of immunology and hygienic inspection. Background technique: [0002] Mercury, commonly known as mercury, is the only metal that exists in a liquid state at room temperature and pressure. It is chemically stable and can evaporate at room temperature. Mercury vapor and mercury compounds are often highly toxic. The use of mercury has a long history, and the ancient people used natural mercury sulfide in the Yin and Shang Dynasties. Mercury has a wide range of uses, and is often used in the manufacture of scientific measuring instruments, medicines, catalysts, mercury vapor lamps, electrodes, thunder mercury, cosmetics, etc. Mercury is ubiquitous in nature, and generally animals and plants contain trace amounts of mercury. my country is a large country in the production, use and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 张海棠范国英王自良葛亚明张慧辉王淑云
Owner HENAN INST OF SCI & TECH
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