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Application of CRISP1 (cystein-rich secretory protein 1) test paper for diagnosing gingival carcinoma

A technology of cysteine ​​and secreted protein, which is applied in the analysis of materials, biological tests, material inspection products, etc., can solve problems such as the inconclusive range of palatal gums, and achieve convenient self-monitoring of the disease, intuitive and reliable judgment of results, and specificity strong effect

Inactive Publication Date: 2015-03-11
SHANGHAI TENTH PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The gingiva on the palatal side of the maxilla is continuous with the palatal mucosa, and there is no clear boundary in clinical practice, so the extent of the palatal gingiva has not yet been determined.

Method used

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  • Application of CRISP1 (cystein-rich secretory protein 1) test paper for diagnosing gingival carcinoma
  • Application of CRISP1 (cystein-rich secretory protein 1) test paper for diagnosing gingival carcinoma
  • Application of CRISP1 (cystein-rich secretory protein 1) test paper for diagnosing gingival carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] The specific embodiments provided by the present invention will be described in detail below in conjunction with the accompanying drawings.

[0012] The reference signs and components involved in the accompanying drawings are as follows:

[0013] 1. Bottom plate 2. Sample pad

[0014] 3. Gold pad 4. Nitrocellulose membrane layer

[0015] 5. Absorbent paper layer 6. Saliva

[0016] 7. Chromatographic direction

[0017] Example 1 Immunohistochemistry

[0018] Experimental materials: normal gingival tissue and gingival cancer tissue; the primary antibody is goat anti-human CRISP1 antibody, and the secondary antibody is donkey anti-goat Dylight594 antibody.

[0019] experimental method:

[0020] Tissue chip is the orderly combination of a large number of tissue samples on the surface of a micro-substrate, with the help of immunohistochemical method for detection.

[0021] 1. Fix the tissue with PFA fixative for 5 minutes, wash with PBS for 10 minutes

[0022] 2. Anti...

Embodiment 2

[0037] Example 2 Enzyme-linked immunosorbent assay (Elisa)

[0038] A total of 90 cases, including 30 cases of gingival cancer (squamous cell carcinoma) case group; 30 cases of other oral diseases (periodontal disease, pulp disease, oral candidiasis) group; 30 cases of healthy control group. The cysteine-rich secreted protein detection kit is used for detection, and the detection steps refer to the instruction manual of the cysteine-rich secreted protein quantitative enzyme-linked detection kit.

[0039] ELISA detection results of CRISP1:

[0040] 1. The gingival cancer group was higher than the other two groups, while there was no difference between the other disease group and the healthy group (P=0.802).

[0041] Table 1 The diagnostic value of saliva CRISP1 in the diagnosis of gingival cancer

[0042] diagnostic index AUC sensitivity specificity false positive rate false negative rate Youden index Urine CRISP1 10 ng / ml 0.829 86.5% 41.3% 60.7...

Embodiment 3

[0045] Example 3 Preparation of gold-labeled antibody complex on test paper

[0046] 1. Colloidal gold preparation

[0047] Add 100ml of 0.005%-0.02% chloroauric acid solution into a round bottom flask and heat to boiling. After boiling, add freshly prepared 0.4%-2% trisodium citrate (Na 3 C 6 h 5 o 7 2H 2 O) 1-2.2ml of aqueous solution, after boiling for 10-25 minutes, continue to stir and cool to room temperature. During this process, it can be seen that the color change of the solution is: golden yellow→black→purple→dark blue→bright red. When the color of the solution completely changes to transparent bright red, the required colloidal gold is obtained. After cooling, put it into a dialysis bag and dialyze it against ultrapure water (1:5000) three times, and finally transfer the dialyzed colloidal gold to a clean glass bottle with a screw cap, and store it in a dark environment at 4°C.

[0048] 2. Connection between gold colloid and protein

[0049] 1. The adsorption...

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Abstract

The invention relates to application of CRISP1 (cystein-rich secretory protein 1) test paper for diagnosing gingival carcinoma in preparation of medical appliances for diagnosing gingival carcinoma. The test paper comprises a substrate, a sample pad, a colloidal gold pad, a nitrocellulose membranous layer and a water-absorption paper layer, wherein the nitrocellulose membranous layer contains a BSA (bovine serum albumin)-saliva CRISP1 coupling substance test line and an anti-mouse secondary antibody quality control line; and the colloidal gold pad contains a colloidal gold labeled anti-saliva CRISP1 antibody compound formed by combining colloidal gold and anti-saliva CRISP1 antibody. The test paper provided by the invention has the advantages of high detection speed, high sensitivity, high specificity, high stability, no need of any instrument or equipment, and visual and reliable judgment result, is simple to operate and easy to master, and is convenient for patients to perform self-monitoring of the state of illness.

Description

technical field [0001] The invention relates to the application of cysteine-rich secreted protein 1 test paper for diagnosing gum cancer. Background technique [0002] Gum cancer (carcinoma of the gingiva) is second only to tongue cancer and buccal cancer among oral cancers, accounting for about 22% of oral cancers. Men outnumber women. More common in 40 to 60 years old. Most of them are squamous cell carcinomas with a high degree of differentiation, with slow growth and no obvious symptoms in the early stage. Ulcer type is more common. Gingival cancer can occur in the gingival mucosa of the lip and buccal side, and can also occur in the gingival mucosa of the tongue and palate. The upper and lower lip and buccal gingival mucosa are adjacent to the buccal mucosa, with the lip and buccal groove as the dividing line. The gingival mucosa on the lingual side of the mandible is adjacent to the floor of the mouth, with the lingual groove as the dividing line. The gingiva on ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/574
Inventor 朱进安万思慧
Owner SHANGHAI TENTH PEOPLES HOSPITAL
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