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Application of SHARPIN gene in preparation of reagent for diagnosing human skin tumors

A skin tumor, a technology for diagnosing people, used in the fields of bioengineering and medicine

Inactive Publication Date: 2016-12-07
梁燕华
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] It has been reported that mutations in the exons of the SHARPIN gene lead to the occurrence of chronic proliferative dermatitis (cpdm) in mice, but there is no international article reporting the relationship between the SHARPIN gene and NMSCs

Method used

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  • Application of SHARPIN gene in preparation of reagent for diagnosing human skin tumors
  • Application of SHARPIN gene in preparation of reagent for diagnosing human skin tumors
  • Application of SHARPIN gene in preparation of reagent for diagnosing human skin tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Preparation of kits for the diagnosis of human skin tumors

[0078] include:

[0079] PCR reaction solution containing Taq enzyme, dNTP, magnesium ion, PCR reaction buffer

[0080] PCR product purification kit Contains a small amount of PCR product purification solution, DNA adsorption column

[0081] Sequencing reaction solution with Big Dye

[0082] Primers, as shown in Table 3 (one of the optional groups):

[0083] table 3

[0084]

[0085]

[0086]Take 3ml of blood from the patient to be tested, and extract DNA from the blood. Dilute the PCR primers in the kit for diagnosing human skin tumors to 1 μmol / μl, and use the extracted DNA as a template to perform a PCR reaction with the provided primers. Use the PCR product purification kit provided by the detection kit to purify the PCR product. The purified product is directly sequenced after the sequencing reaction. Observe whether there are mutations in the sequence obtained by sequencing.

Embodiment 2

[0088] The expression of SHARPIN protein in NMSC was detected by immunohistochemical method:

[0089] Preparation Kit

[0090] 1. The kit contains reagents

[0091] Rabbit anti-human SHARPIN antibody (100 μg IgG / mL), rabbit anti-immunohistochemical kit (reagent A: blocking solution is 10% goat serum, reagent B: anti-rabbit biotinylated secondary antibody, reagent C: streptavidin-labeled HRP, reagent D: 20 times concentrated DAB substrate buffer, reagent E: 20 times concentrated DAB chromogenic solution, reagent F: 20 times concentrated hydrogen peroxide solution).

[0092] 2. Specific implementation steps

[0093] 1. Making wax blocks and HE staining

[0094] Under local anesthesia, a small piece of skin from the patient's skin lesion was surgically excised, and the specimen was fixed in 10% formaldehyde solution as soon as possible, and the excised skin specimen was further processed according to the following steps. The whole production process is as follows: skin specim...

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Abstract

The invention discloses application of SHARPIN gene in the preparation of a reagent for diagnosing human skin tumors. Genome DNA sequence of the SHARPIN gene is as shown in SEQ ID NO. 1, a protein sequence is as shown in SEQ ID NO. 2, and SHARPIN gene mutation site is at c.103 with protein sequence mutation site at p.35, or SHARPIN gene mutation site is at c.329 with protein sequence mutation site at p.110, or SHARPIN gene mutation site is at c.214 with protein sequence mutation site at p.72, or SHARPIN gene mutation site is at c.221 with protein sequence mutation site at p.74, or SHARPIN gene mutation site at c.347 with protein sequence mutation site at p.116. The technical problem to be solved is to provide a method to assist in diagnosing NMSC (non-melanoma skin cancer) and application of SHARPIN GENE kit in the preparation of reagents for diagnosing human skin tumors.

Description

technical field [0001] The invention relates to the fields of bioengineering and medicine, in particular to the use of a SHARPIN gene in preparing reagents for diagnosing human skin tumors. Background technique [0002] Both squamous cell carcinoma (SCC) and basal cell carcinoma (BCC) belong to nonmelanoma skin cancer (NMSC), and both are common skin malignancies in humans. Like other malignant tumors, once tissue cells are transformed into malignant cells, their biological behavior will also change accordingly, showing continuous and non-self-limited proliferation, and finally forming tumors. Their formation is a multi-step and multi-factor process. , the traditional concept is that it is the result of the accumulation of a gene mutation, involving important molecular events such as oncogene mutation, tumor suppressor gene inactivation, or apoptosis regulatory gene abnormality. [0003] Skin SCC is a malignant tumor originating from epidermis or adnexal keratinocytes. Alth...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574
CPCC12Q1/6886C12Q2600/156G01N33/5743G01N33/68G01N2333/47
Inventor 梁燕华
Owner 梁燕华
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