Method and kit for prognosis and guiding treatment of patients with laryngeal papilloma

A kit, HLA-DRB1 technology, applied in biochemical equipment and methods, microbial measurement/testing, DNA/RNA fragments, etc., can solve difficult problems and achieve simple operation, great application value, and low cost Effect

Active Publication Date: 2018-03-23
BEIJING CHILDRENS HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is very difficult to design specific PCR amplification primers for the HLA-DRB1*03:01 genotype

Method used

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  • Method and kit for prognosis and guiding treatment of patients with laryngeal papilloma
  • Method and kit for prognosis and guiding treatment of patients with laryngeal papilloma
  • Method and kit for prognosis and guiding treatment of patients with laryngeal papilloma

Examples

Experimental program
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Effect test

Embodiment 1

[0035] The primers of the invention are used to perform qualitative detection of HLA-DRB1*03 low-resolution type on human blood samples. Specific steps are as follows:

[0036] 1) Extraction of peripheral blood genomic DNA

[0037] The peripheral blood samples of 6 volunteers (sample numbers 14, 15, 19, 21, 22, 23) with known HLA-DRB1 genotypes (DRB1*03 low-resolution type were positive in 3 cases and negative in 3 cases) were collected by sequencing. 1 ml of each blood was extracted using the blood genomic DNA extraction kit from Promega Company, and was extracted according to the operating instructions of the kit.

[0038] 2) PCR amplification

[0039] Different primers are used to carry out PCR amplification reaction on the DNA extracted in step 1). Use SEQ ID No.1 (GTT TCT TGG AGT ACT CTA CGT CTG) and SEQ ID No.2 (TCC ACC CGG CCC CGC TTC TG) as HLA-DRB1*03 low resolution qualitative primer pair, SEQ ID No.3 ( TTC TTC AAT GGG ACG GAG CG) and SEQID No.4 (GGT CCT TCT GGC ...

Embodiment 2

[0050] Using the primers of the present invention, perform qualitative detection of HLA-DRB1*03 low-resolution type in RRP patients, and further conduct sequencing analysis. Specific steps are as follows:

[0051] Select 20 RRP patients, adopt the method described in Example 1, extract the peripheral blood genomic DNA of the subjects, perform PCR amplification reaction, and perform 2% agarose gel electrophoresis on the results of the amplification reaction, and analyze the Qualitative results of HLA-DRB1*03 low-resolution type of the test subject. At the same time, for the qualitative positive results of HLA-DRB1*03 low-resolution type, the corresponding PCR amplification product was sent to Shenggong Bioengineering (Shanghai) Co., Ltd. for sequencing, and compared with the sequence published on the IMGT website to analyze the corresponding high-resolution subtypes. In addition, the peripheral blood genomes of the 20 RRP patients were simultaneously sent to Beijing Bofurui G...

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Abstract

The invention relates to a gene detection method and kit capable of being used for clinical detection. The method is aimed at HLA-DRB1*03 low-resolution gene and can realize accurate, rapid, simple and inexpensive detection, and can be used for providing important information for treatment and prognosis of clinically relevant diseases such as recurrent laryngeal papilloma and the like.

Description

technical field [0001] The invention belongs to the field of gene diagnosis, and relates to a gene detection method and kit, as well as the application in clinical related diseases. technical background [0002] Recurrent respiratory papilloma (RRP) is the most common benign tumor of the laryngeal epithelial tissue, and the main pathogens are human papillomavirus 6 and 11 subtypes. Repeated resection is the main treatment for this disease, but it cannot prevent recurrence. During the active stage of the disease, most patients may need to undergo an operation every 2-3 months, and some children may even undergo hundreds of operations in their lifetime, which brings a heavy burden to the patients and their families. In addition, the frequent recurrence of RRP increases the possibility of spreading the lesions to the lower respiratory tract, which in turn makes the disease more aggressive. [0003] At present, there is no effective cure for RRP, but a variety of drugs have be...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/118C12Q2600/156
Inventor 倪鑫王国亮邰隽张杰吴逊尧桂晋刚赵靖王华王桂香王生才唐力行
Owner BEIJING CHILDRENS HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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