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A primer set, reagent and/or kit, system and application thereof for detecting lung cancer chemotherapy-related genes

A technology for detecting reagents and primer sets, which is applied in the field of primer sets for detecting lung cancer chemotherapy-related genes, can solve the problems of inability to detect unknown mutations, low throughput of in situ hybridization, high probe prices, etc. Ease of analysis and high sensitivity

Active Publication Date: 2020-06-23
SIMCERE DIAGNOSTICS CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the PCR-direct sequencing method has low throughput and consumes a lot of samples, and the average detection of each site needs to consume 100ng; the accuracy of the PCR-pyrosequencing method for sequences with multiple single-base repeats is low; fluorescence quantitative The PCR method cannot detect unknown mutations, and the probes are expensive; PCR-gene chip method and PCR-electrophoresis analysis method are also difficult to detect unknown mutations; the throughput of in situ hybridization method is too low, and the workload is heavy when a large number of typing , and only applicable to some SNP typing
[0004] Among these detection methods, there are few existing technologies that can be applied to the accurate genome detection of lung cancer chemotherapy drugs effectively and at low cost. The detection means of chemotherapeutic drug genome, especially propose the present invention

Method used

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  • A primer set, reagent and/or kit, system and application thereof for detecting lung cancer chemotherapy-related genes
  • A primer set, reagent and/or kit, system and application thereof for detecting lung cancer chemotherapy-related genes
  • A primer set, reagent and/or kit, system and application thereof for detecting lung cancer chemotherapy-related genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Primer Design Optimization

[0048] 1) Primer sequence optimization

[0049] According to the site in Table 1, the present invention comprehensively designs PCR primers and single-base extension primers based on software and design experience, uses gDNA as a template to detect the working efficiency of primers, and finally screens out the best PCR primers and single-base extension primers through multiple rounds of design and repeated optimization. For base extension primers, see the primer sequences in Tables 2 and 3 for details.

[0050] Table 2. PCR primer sequences

[0051]

[0052]

[0053] Table 3. Single-point extension primer sequences

[0054]

[0055]

[0056] 2) Optimization of annealing temperature system

[0057] The annealing temperature is an important parameter in the reaction system. During the experiment, it was found that when the annealing temperature of the PCR reaction was set at 56°C as usual, the amplification efficiency ...

Embodiment 2

[0058] Embodiment 2 system verification

[0059] The verification of the system of the present invention includes accuracy, specificity, sensitivity, precision and comparison between personnel, etc., specifically:

[0060] The accuracy verification scheme of the present invention: each site detects 20 cases, compared with Sanger sequencing, the expected target is 95%.

[0061] The specificity verification scheme of the present invention: included in the accuracy, the expected target is 95%.

[0062] The sensitive verification scheme of the present invention: included in the accuracy, the expected target is 95%.

[0063] The precision verification scheme of the present invention (including intra-batch, inter-batch, and personnel comparison, not involving inter-instrument comparison) has an expected target of 95%.

[0064] Intra-assay precision: each sample was repeated 3 times in the same batch, and the intra-assay precision was compared:

[0065] Batch-to-batch precision: t...

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Abstract

The invention relates to a primer group, reagent and / or kit and system for detecting lung cancer chemotherapy related genes, and application. The primer group contains primers for detecting gene mutation of ERCC1, MTHFR, GSTP1, XRCC1, DYNC2H1, ABCB1, CYP2C8*3, TP53, NQO1, CBR3, SOD2, CYP2C19, UGT1A1*6, TYMS, NT5C2 and CDA. The primer group, the reagent and / or the kit, the system and the application have the advantages of high accuracy, high specificity, high sensitivity, good precision and the like, and meanwhile, further have the advantages of sample saving, short detection period, easy operation, convenient analysis and the like.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a primer set, reagent and / or kit, system and application thereof for detecting lung cancer chemotherapy-related genes. Background technique [0002] Chemotherapy is often accompanied by various adverse reactions. For example, bone marrow toxicity is a very common non-specific toxicity of chemotherapy, manifested as leukopenia, thrombocytopenia (more common with gemcitabine and other drugs), chemotherapy-related anemia (more common with cisplatin and other drugs); Chemotherapy-related vomiting, cisplatin, high-dose cyclophosphamide (≥1000mg / m2) and other drugs, the incidence of vomiting is as high as 90% to 100%; chemotherapy-related diarrhea, in mild cases, can reduce the quality of life of patients, and in severe cases can cause Acid-base imbalance, water-electrolyte disturbance and other complications occur, even leading to death of the patient. Diarrhea is the main dose-limiting ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/106C12Q2600/156
Inventor 陈珊珊赵妍程江月李杜衡任用
Owner SIMCERE DIAGNOSTICS CO LTD
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