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Application of specific primers of clc gene in preparation of kit for detection of subtypes of chronic sinusitis with nasal polyps

A specific technology for chronic sinusitis, applied in the field of biomedicine, can solve the problems of different number of inflammatory cells, difficulty in batch operation, fear of patients, etc., to achieve the effect of high identification accuracy, saving labor costs, and solving human errors

Active Publication Date: 2021-03-23
张罗 +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of pathological biopsy of nasal mucosa are as follows: 1. It is an invasive examination: it increases the risk of infection for patients, and is not suitable for people with low immunity such as children and the elderly; nasal cavity bleeding often causes fear and worry in patients
2. It is difficult to obtain real-time dynamic change information of the disease: pathological biopsy of the healing mucosa cannot be performed after the operation
However, clinical data show that the pathological classification of chronic sinusitis with nasal polyps will vary with the outcome of drug treatment, surgical treatment, etc. Pathological biopsy results before treatment cannot represent the characteristics of all disease stages
3. Time-consuming and increase the cost of medical treatment, usually 3-4 working days from obtaining tissue samples to obtaining pathological results
Due to the inability to obtain the results on the same day or the next day, extra transportation, accommodation, and registration fees will be incurred for out-of-town patients seeking medical treatment, which increases the cost of medical treatment
4. There are certain human errors, and the number of inflammatory cells counted by different pathologists may be different, which affects the judgment of polyp typing
5. Histopathological sections are relatively limited, and can only reflect the inflammatory state of the specimen at the section position, but cannot reflect the whole picture of the tissue, which may cause misdiagnosis
6. Each film needs to be manually counted by pathologists, which is difficult to operate in batches

Method used

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  • Application of specific primers of clc gene in preparation of kit for detection of subtypes of chronic sinusitis with nasal polyps
  • Application of specific primers of clc gene in preparation of kit for detection of subtypes of chronic sinusitis with nasal polyps
  • Application of specific primers of clc gene in preparation of kit for detection of subtypes of chronic sinusitis with nasal polyps

Examples

Experimental program
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Effect test

Embodiment 1

[0068] A kit for detecting subtypes of chronic sinusitis with nasal polyps, comprising the following reagents:

[0069] Reagents for extracting RNA from nasal polyp tissue: 20mL RNA extraction solution Trizol or RNAiso Blood or RNAiso Plus or others containing phenol, guanidine isothiocyanate, 8-hydroxyquinoline, guanidine isothiocyanate, β-mercaptoethanol, etc. Substances that can rapidly disrupt cells and inhibit nucleases released from cells; 2mL of chloroform; 20mL of isopropanol; 40mL of 65-90% ethanol; 5mL of RNase- and DNase-free water;

[0070] Reagents for reverse transcription of extracted RNA into cDNA: 40 μL of reverse transcription mixture (containing enzymes required for reverse transcription, RNase inhibitors, random 6-nucleotide primers, polythymine, T repeat oligonucleotides , deoxyribonucleotide triphosphate mixture, buffer, etc.), 160 μL of RNase- and DNase-free water; the RNase- and DNase-free water is used to make up the system, dissolve, and dilute RNA; ...

Embodiment 2

[0073] A kit for detecting subtypes of chronic sinusitis with nasal polyps, comprising the following reagents:

[0074] Reagents for extracting RNA from nasal polyp tissue: 1mL RNA extraction solution Trizol or RNAiso Blood or RNAisoPlus or other substances containing phenol, guanidine isothiocyanate, 8-hydroxyquinoline, guanidine isothiocyanate, β-mercaptoethanol, etc. Substances that can rapidly disrupt cells and inhibit nucleases released from cells; 0.2mL of chloroform; 0.2mL of isopropanol; 0.2mL of 65-90% ethanol; 0.05mL of RNase- and DNase-free water;

[0075] Reagents for reverse transcription of extracted RNA into cDNA: 2 μL of reverse transcription mixture (containing enzymes required for reverse transcription, RNase inhibitors, random 6-nucleotide primers, polythymine, T repeat oligonucleotides , deoxyribonucleotide triphosphate mixture, buffer, etc.), 7 μL of RNase- and DNase-free water; the RNase- and DNase-free water is used to make up the system, dissolve, and d...

Embodiment 3

[0078] A kit for detecting subtypes of chronic sinusitis with nasal polyps, comprising the following reagents:

[0079] Reagents for extracting RNA from nasal polyp tissue: 0.1mL RNA extraction solution Trizol or RNAiso Blood or RNAiso Plus or other containing phenol, guanidine isothiocyanate, 8-hydroxyquinoline, guanidine isothiocyanate, β-mercaptoethanol 0.05mL of chloroform; 0.015mL of isopropanol; 0.0075mL of 65-90% ethanol; 0.01mL of RNase- and DNase-free water ;

[0080] Reagents for reverse transcription of total RNA into cDNA: 1 μL of reverse transcription mixture (containing enzymes required for reverse transcription, RNase inhibitors, random 6-nucleotide primers, polythymine, T repeat oligonucleotides, Deoxyribonucleotide triphosphate mixture, buffer, etc.), 0-10 μL of RNase- and DNase-free water; the RNase- and DNase-free water is used to make up the system, dissolve, and dilute RNA;

[0081] Reagents for real-time fluorescent quantitative PCR reaction of CLC gene...

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Abstract

The invention provides a kit for detecting chronic sinusitis with subtype nasal polyp as well as an application of a CLC gene as a biomarker. The kit comprises a specific primer of CLC gene. The CLC gene can be used for preparing a product for detecting the chronic sinusitis with subtype nasal polyp as a biomarker. According to the kit, the CLC gene is screened to be used as the biomarker by virtue of proteomics and transcriptomics methods, so that a method for detecting the chronic sinusitis with subtype nasal polyp can be realized by adopting the kit, and the subtype nasal polyp can be rapidly identified; compared with the traditional pathological examination method, the consumed time is shorter; and the kit can simultaneously massively and rapidly detect samples, so that the manpower cost and the medical treatment cost can be reduced. More importantly, the characteristics of histopathology can be comprehensively reflected.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of a CLC gene-specific primer in preparing a kit for detecting subtypes of chronic sinusitis with nasal polyps. Background technique [0002] Chronic rhinosinusitis with nasal polyps (CRSwNP) is a chronic inflammation of the sinus mucosa, and physical examination shows the formation of polyps in the nasal cavity or middle nasal passage. Common symptoms of CRSwNP are nasal congestion, runny nose or backflow, hyposmia, facial fullness or pressure, and last for more than 12 weeks. The prevalence rate is about 0.5-4%. CRSwNP is often accompanied by asthma and allergic rhinitis. It has been reported that 7% of asthmatic patients suffer from CRSwNP, while 26-48% of CRSwNP suffer from asthma. The pathogenesis of CRSwNP is still uncertain. The destruction of mucosal epithelial cells, the imbalance of host immune system and the invasion of pathogenic micro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/6851
CPCC12Q1/6851C12Q1/6883C12Q2600/112C12Q2600/158C12Q2600/166C12Q2531/113C12Q2545/101
Inventor 张罗王成硕闫冰刘畅
Owner 张罗
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