Fast detection test paper tape of lead ion aurosol immune layer as well as preparation method and application thereof

A detection test paper, immunochromatography technology, applied in the field of immunology, to achieve the effect of low cost, simple operation, and guaranteed benefits

Active Publication Date: 2010-02-24
广东曜城科技园管理有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report on the rapid detection method of lead ion colloidal gold immunochromatography at home and abroad, and there is no such test strip product in the domestic ...

Method used

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  • Fast detection test paper tape of lead ion aurosol immune layer as well as preparation method and application thereof
  • Fast detection test paper tape of lead ion aurosol immune layer as well as preparation method and application thereof
  • Fast detection test paper tape of lead ion aurosol immune layer as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] (1) Preparation and purification of anti-lead ion monoclonal antibody: use bifunctional diethylenetriaminepentaacetic acid (p-SCN-Bn-DTPA) chelating agent to couple lead ions to bovine serum albumin (BSA), mix BALB / c mice were immunized with incomplete adjuvant, the splenocytes of the immunized mice were fused with SP2 / 0 myeloma cells, and positive cell lines stably secreting anti-lead ion monoclonal antibody (MAb-Pb) were screened out and expanded for culture. Inject cells into mice to induce ascites; take 1 volume of ascites, add 2 volumes of 0.06mol / L sodium acetate buffer solution with pH5.0 (59ml sodium acetate solution with a concentration of 0.06mol / L + 41ml with a concentration of 0.06mol / L L of glacial acetic acid mixed), then adjust the pH to 4.5 with 1mol / L hydrochloric acid; slowly add octanoic acid with a volume ratio of 33ul / ml dropwise within 30min under stirring at room temperature, let stand at 4°C for 2h, then 4°C Centrifuge at 10000rpm / min for 30min t...

Embodiment 2

[0055] To verify the detection specificity test of the present invention:

[0056] The lead ion colloidal gold immunochromatographic rapid detection test strip that following heavy metal utilization embodiment makes carries out specificity cross reaction test, and the result is as follows:

[0057] Table 1 Cross-reactivity of other heavy metal lead

[0058]

[0059] The results showed that the antibody except for Pd 2+ Besides, there is almost no cross-reaction to several other metals, so this colloidal gold test strip can be used to detect Pd 2+ .

Embodiment 3

[0061] Verify reliability test of the present invention: (see step b and c image 3 shown, step d see Figure 4 shown)

[0062] a. Prepare Pd first 2+ A series of gradient standard solutions (10, 20, 50, 80, 100 ng / mL) each 50 μL, and then add 50 μL 1mmol / L DTPA solution as a chelating agent, mix well, so that lead ions and DTPA are fully chelated into Pd-DTPA, as The sample solution to be tested.

[0063] b. Use a dropper to draw 80-100 μL of the sample solution to be tested obtained in step a, drop it on the sample pad of the test strip, and start timing after dropping the sample;

[0064] c. Read the result 3 to 5 minutes after adding the sample. When reading, place the test strip vertically in front of the observer with one end of the sample pad facing down;

[0065] d. Judgment of results: C line is displayed as a red line. When the T line develops color, the result is negative, and the lead ion concentration in the sample to be tested is less than 50ng / ml; when the T...

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Abstract

The invention discloses a fast detection test paper tape a lead ion aurosol immune layer as well as a preparation method and the application thereof. The preparation method of the test paper tape comprises the following steps: (1) preparing and purifying a lead ion resistant monoclonal antibody; (2) preparing a detection antigen Pb-DTPA-OVA; (3) preparing an aurosol solution; (4) preparing an aurosol solution marked with the lead ion resistant monoclonal antibody; (5) spraying metal; (6) enveloping C and a T wire on a nitrocellulose membrane; and (7) assembling the test paper tape. A method detecting lead ions by using the test paper tape comprises the following steps: a. adding a DTPA chelating agent into a sample digestion solution to be detected so that the lead ions and the DTPA chelating agent are sufficiently chelated; b. using a burette to absorb the sample digestion solution to be detected, dripping 3 drips of the sample digestion solution on a sample pad of the test paper tape, and starting to time after sample addition; c. reading a result 3-5 minutes after the sample addition; when reading the result, taking the test paper tape to be perpendicular to the front face of an observer in such a way that one end of the sample pad is downward; and d. judging the result.

Description

technical field [0001] The invention relates to a rapid detection test strip of lead ion colloidal gold immunochromatography and its preparation method and application, belonging to the technical field of immunology. Background technique [0002] Lead (lead) atomic number 82 is one of the heavy metals that seriously endanger human health. Among all known toxic substances, lead is the most recorded in the book. Lead pollution comes from production wastewater and wastes such as mining, smelting, rubber production, dyes, printing, ceramics, lead glass, solder, cables and lead pipes. In addition, tetraethyl lead in automobile exhaust is a highly toxic substance. With the rapid development of industry and agriculture in our country, the pollution of heavy metals in the environment is becoming more and more serious. The poison caused by heavy metal pollution persists and will enter the food chain with the recirculation of soil and water, posing a threat to food safety and endang...

Claims

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Application Information

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IPC IPC(8): G01N33/558G01N33/577G01N33/532G01N33/52C07K16/44C07K14/77C07K1/113
Inventor 唐勇向军俭翟一凡邓宁王宏杨红宇
Owner 广东曜城科技园管理有限公司
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