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Colloidal gold immunochromatography kit for quickly detecting ovarian cancer tumor marker Legumain and preparation method of kit

A tumor marker, immunochromatography technology, applied in biological tests, measurement devices, analytical materials, etc., can solve problems such as obstacles to the popularization and promotion of early detection of ovarian cancer, high requirements for equipment and operators, and low diagnostic specificity of ovarian cancer. problems, to achieve the effect of small sample size, elimination of false positives, and mature preparation process

Inactive Publication Date: 2020-05-01
南京拂晓生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these methods can accurately diagnose ovarian cancer when they are used in conjunction with each other, they have high requirements for equipment and operators, which increases the cost of detection and hinders the popularization and promotion of early detection of ovarian cancer.
[0004] The detection of tumor markers is widely used in the early screening of tumors. Carbohydrate antigen 125 (carbohydrateantigen125, CA125) is a commonly used tumor marker in the diagnosis of ovarian cancer, but its specificity is low when it is used in the diagnosis of ovarian cancer, and it is easy to appear. False positives and other problems, so it is necessary to find new tumor markers to replace it or complement it

Method used

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  • Colloidal gold immunochromatography kit for quickly detecting ovarian cancer tumor marker Legumain and preparation method of kit
  • Colloidal gold immunochromatography kit for quickly detecting ovarian cancer tumor marker Legumain and preparation method of kit
  • Colloidal gold immunochromatography kit for quickly detecting ovarian cancer tumor marker Legumain and preparation method of kit

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Experimental program
Comparison scheme
Effect test

Embodiment

[0043] The present invention will be described below through specific embodiments, but the protection scope of the present invention is not limited to the following examples.

[0044] The Anti-Legumain-Ab1 and Anti-Legumain-Ab2 used in the kit of the present invention were purchased from Abcom Company, and the product numbers are ab183028 and ab232870 in sequence.

Embodiment 1

[0045] Example 1. Preparation of Colloidal Gold Kit for Rapid Detection of Legumain Anti-Legumain-Ab1 and Anti-Legumain-Ab2 are monoclonal antibodies against different epitopes of Legumain respectively.

[0046] (1) Preparation of colloidal gold

[0047] Take 80mL of ultrapure water in a clean Erlenmeyer flask and put it into a clean rotor, place it on a heatable magnetic stirrer and heat until bubbles come out evenly, then quickly add 1mL of 1% chloroauric acid solution, and heat to boiling Immediately add 1mL of 1.5% trisodium citrate, at this time the color of the solution in the Erlenmeyer flask changes from grayish black to wine red, continue heating for 7min, remove the Erlenmeyer flask, let it cool naturally at room temperature, and dilute to 100mL with a volumetric flask. Sealed for later use.

[0048] (2) Antibody labeling

[0049] Take 1 mL of the colloidal gold prepared in step (1), add potassium carbonate solution with a concentration of 0.1 mol / L, and adjust the...

Embodiment 2

[0064] Embodiment 2, the method for judging the result of the colloidal gold-labeled anti-Legumain antibody kit

[0065] The result evaluation method of Legumain rapid detection kit is as follows: figure 2 As shown, when both the C quality control band 1 and the T detection band 2 are colored, the detected sample contains Legumain above the sensitivity (corresponding to figure 2 Middle 3); when the C quality control band 1 develops color, but the T detection band 2 does not develop color, it means that the tested sample does not contain or contain Legumain below the sensitivity (corresponding to figure 2 Middle 4); if the C quality control band 1 does not develop color, no matter whether the T detection band 2 develops color or not, the detection effect is invalid (corresponding to figure 2 Middle 5).

[0066] Performance testing experiment of Legumain colloidal gold rapid detection kit:

[0067] 1. Sensitivity test Dilute the Legumain standard with negative human serum...

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Abstract

The present invention discloses a colloidal gold immunochromatography kit for quickly detecting an ovarian cancer tumor marker Legumain and a preparation method of the kit. The kit comprises a PVC bottom plate, a sample pad, a combination pad, a reaction pad and a water absorption pad, wherein the PVC bottom plate is rectangular, the sample pad and the water absorption pad are symmetrically laid on a group of the short edges of the rectangular PVC bottom plate, the reaction pad is arranged between the sample pad and the water absorption pad, and the combination pad is further arranged betweenthe sample pad and the reaction pad; and the reaction pad is provided with a colloidal gold labelled mouse anti-Legumain antibody coated detection line and a goat anti-mouse IgG coated quality controlline. Colloidal gold as a lateral chromatography kit label is mature in preparation process and stable in performance; the specificity of a detection result can be enhanced and false positive in a detection process can be eliminated, so that the detection result is more accurate; and the amount of samples which need to be detected is small, and the kit is applicable to ovarian cancer screening, household self-examination and bedside rapid detection of vast women.

Description

technical field [0001] The invention relates to the field of biological immunochromatography detection methods, in particular to a colloidal gold immunochromatography kit for rapid detection of ovarian cancer tumor marker Legumain and a preparation method thereof. Background technique [0002] Ovarian cancer is a common malignant tumor in gynecology, and its mortality rate ranks first among all gynecological malignant tumors, endangering women's health. Because ovarian cancer usually has no obvious symptoms in the early stage and lacks simple and effective early diagnosis methods, about 70% of the patients are already in the advanced stage when they come to the doctor due to symptoms, and the 5-year survival rate of the advanced stage patients is 20%-30%, while the early stage patients The 5-year survival rate is 70-90%. Therefore, early detection of ovarian cancer plays an important role in the selection of treatment options and in improving the prognosis of patients. [...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/574G01N33/559
CPCG01N33/68G01N33/57449G01N33/57484G01N33/559
Inventor 李伟赵树杰
Owner 南京拂晓生物科技有限公司
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