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Preparation method of lung cancer marker detection immunochromatographic test paper

A technology of immunochromatographic test strips and markers, which is applied in the direction of measuring devices, analysis materials, instruments, etc., can solve the problems that the sensitivity and specificity of tumor markers are difficult to meet the requirements of early diagnosis, differential diagnosis, curative effect and prognosis evaluation, and achieve High sensitivity, easy test operation and accurate test results

Inactive Publication Date: 2012-11-07
SHANGHAI NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the sensitivity and specificity of the detection of a single tumor marker is difficult to meet the clinical requirements for early diagnosis, differential diagnosis, curative effect and prognosis evaluation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Preparation of colloidal gold-neuron-specific enolase ((NSE)) antibody complex:

[0059] Utilize the colloidal gold of newly prepared 40nm to connect with (NSE) antibody, concrete implementation steps are as follows:

[0060] 1. Take 1mL colloidal gold solution, add 10uL 0.1M K 2 CO 3 Adjust the pH of the solution to 9.0 and mix well.

[0061] 2. Add 8uL (NSE) monoclonal antibody (L1C00501) into the colloidal gold solution, mix well under vortex, and let stand for 10min.

[0062] 3. Add 200uL 10% BSA solution, mix well, and let stand for 10min.

[0063] 4. Centrifuge the colloidal gold solution obtained above at 10,000 rpm for 15 min under refrigeration.

[0064] 5. Aspirate the supernatant, redisperse the precipitate with 1 mL borate buffer solution (containing 0.5-1.0% sucrose, 2-5% BSA, 0.01M boric acid), and then centrifuge once at high speed.

[0065] 6. Aspirate the supernatant, concentrate to 1 / 10 of the original volume, and store at 4°C.

Embodiment 2

[0067] Preparation of colloidal gold-carcinoembryonic antigen ((CEA)) antibody complex:

[0068] Utilize the colloidal gold of newly prepared 40nm to connect with (CEA) antibody, concrete implementation steps are as follows:

[0069] 1. Take 1mL colloidal gold solution, add 10uL 0.1M K 2 CO 3 Adjust the pH of the solution to 9.0 and mix well;

[0070] 2. Add 10uL (CEA) monoclonal antibody (L1C00202) into the colloidal gold solution, mix well under vortex, and let stand for 10min;

[0071] 3. Add 200uL 10% BSA (bovine serum albumin) solution, mix well, and let stand for 10min;

[0072] 4. Centrifuge the colloidal gold solution obtained above at 10,000 rpm for 15 min under refrigeration;

[0073] 5. Aspirate the supernatant, redisperse the precipitate with 1mL borate buffer solution (containing 0.5-1.0% sucrose, 2-5% BSA, 0.01M boric acid), and then centrifuge once at high speed;

[0074] 6. Aspirate the supernatant, concentrate to 1 / 10 of the original volume, and store at ...

Embodiment 3

[0076] Preparation of double antibody sandwich (NSE), (CEA) test strips:

[0077] 1. Preparation of the sample pad: select glass cellulose membrane as the sample pad material, cut into strips of 5.0×30.0 cm in size, put it into the sample pad blocking solution 0.01mol / L PBS (phosphate buffer solution) (PH=7.4 ), 1.0-2.0% BSA (bovine serum albumin); 2.0-2.0% sucrose, 0.1-0.5% Tween, 0.1-1.0% PVP (polyvinylpyrrolidone)) for 30 minutes, and dried at 37°C for later use.

[0078] 2. Preparation of conjugation pads: glass cellulose membrane was selected as the conjugation pads, and cut into strips with a size of 1.0×30.0 cm for later use.

[0079] 3. Preparation of NC (nitrocellulose membrane): stick the NC membrane on the bottom plate, and draw (CEA), (NSE) monoclonal antibody and goat anti-mouse IgG antibody on different positions on the NC membrane with a scriber, as a detection tape and quality control tape.

[0080] 4. Assembly of immunochromatography test strips: Paste absor...

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Abstract

The invention belongs to the cancer detection technology, relating to a lung cancer marker detection immunochromatographic test paper and an application. The detection of a single tumour marker in the prior art can not meet the requirements of clinical early diagnosis and differential diagnosis. The invention realizes the early diagnosis of lung cancer by jointly detecting NSE and CEA in serum with a double antibody sandwich immunochromatography. The invention comprises the following preparation steps: preparing aurosol-neure NSE and aurosol-carcinoembryonic antigen CEA antibody compounds; preparing double antibody sandwich NSE and CEA detection test strips. The invention comprises the following steps in lung cancer detection: configurating series concentration NSE and CEA standard substances; preparing NSE and CEA mixed antigen solution with serum of normal people; inserting the sample pad ends of the test strips into the above concentration series standard substances respectively toobserve the changes in colours of T and C lines on a NC film. The invention has the advantages of convenient detection operation, high sensitivity and precise result, and can determine lung cancer recurrence 4-12 weeks ahead.

Description

technical field [0001] The invention belongs to cancer detection technology, in particular to an immunochromatographic test paper for detection of lung cancer markers and its application. Background technique [0002] Lung cancer is the most common malignant tumor with the highest morbidity and mortality rate and poor treatment effect. Because there are no specific clinical symptoms and manifestations in the early stage of the disease, most of the patients are already in the late stage when they have clinical manifestations, and the clinical treatment effect and median survival time are not satisfactory. Therefore, early detection and diagnosis of lung cancer is of great significance to the treatment and prognosis of lung cancer. [0003] Neuron-specific enolase (NSE) is an acidic protease unique to neurons and neuroendocrine cells. The content in lung cancer tissue is 3-35 times that in normal lung tissue; The most sensitive and specific tumor marker; it is also a tumor m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/574G01N33/532
Inventor 沈鹤柏孙红英方菲赵露晶
Owner SHANGHAI NORMAL UNIVERSITY
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