PCR kit for detecting human NRAS gene mutation

A kit and human detection technology, applied in the field of PCR kits for detection of human NRAS gene mutations, can solve the problems of long detection cycle and low detection sensitivity

Inactive Publication Date: 2020-12-25
上海佰臻生物科技有限公司
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  • Abstract
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  • Claims
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Problems solved by technology

After decades of clinical application, it has been recognized by the majority of medical staff, but the detection sensitivity of this technology is relatively low, and the detection cycle is long

Method used

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  • PCR kit for detecting human NRAS gene mutation
  • PCR kit for detecting human NRAS gene mutation
  • PCR kit for detecting human NRAS gene mutation

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Embodiment 1

[0052] Such as Figure 1-5 As shown, the present invention provides a PCR kit for detecting human NRAS gene mutation. The kit includes nucleic acid amplification reagents and reference substances, wherein the nucleic acid amplification reagents include NRAS Q61H reaction solution, NRAS Q61K reaction solution, NRASQ61L reaction solution, NRAS Q61R reaction solution, the reaction solution contains specific primers and probes for detecting NRAS Q61H, Q61K, Q61L, Q61R mutations, each reaction solution contains a pair of upstream primers and downstream primers, and a TaqMan fluorescent probe. Primer probe sequences and modifications are as follows:

[0053] Q61H reaction solution: upstream primer 5'-GATTTTGGTCTAGCTATAAA -3'

[0054] Downstream primer 5'-CACAAAATGGATCCAGAC-3'

[0055] Fluorescent probe 5’-FAM-GAAATCTCGATGGAGTGGGTCC-TAMRA-3’

[0056] Q61K reaction solution: upstream primer 5'-GATTTTGGTCTAGCTATAAA -3'

[0057] Downstream primer 5'-CACAAAATGGATCCAGAC-3'

[0058] F...

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Abstract

The invention discloses a PCR kit for detecting human NRAS gene mutation. The kit comprises a nucleic acid amplification reagent and a reference substance, wherein the nucleic acid amplification reagent comprises an NRAS Q61H reaction solution, an NRAS Q61K reaction solution, an NRAS Q61L reaction solution and an NRAS Q61R reaction solution, the reaction solutions respectively contain specific primers and probes for detecting of NRAS Q61H, Q61K, Q61L and Q61R mutations, and each reaction solution contains a pair of upper primer and lower primer, and a TaqMan fluorescent probe. The kit adopts an ARMS-PCR method, and the specific primers and probes aiming at Q61H, Q61K, Q61L and Q61R sites are respectively designed. The kit can be used for detecting mutations of Q61H, Q61K, Q61L and Q61R sites of NRAS genes of tumor tissue DNA samples of patients suffering from colorectal cancer, thyroid cancer and melanoma. The kit can be used for detecting mutations as low as 0.1% in the total amount of 50ng DNA.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a PCR kit for detecting human NRAS gene mutation. Background technique [0002] NRAS gene is one of the most important proto-oncogenes in human beings. It is located on human chromosome 1p13.2 and consists of 7 exons, encoding a protein of about 21kDa. The NRAS mutation is mainly located in the 61st amino acid coding subunit of exon 3. This mutation will affect the GTP hydrolysis mediated by RAS, and will cause the NRAS protein to maintain an active state, no longer dependent on the stimulation of the superior signal, and continue to bind to GTP. state, resulting in abnormally active downstream signaling pathways, thereby promoting cell growth and proliferation, cell proliferation out of control and cancerous. [0003] Colorectal cancer (CRC) is a common malignant tumor. Studies have shown that the occurrence of CRC is closely related to a variety of gene mutations, including KRAS...

Claims

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Application Information

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IPC IPC(8): C12Q1/6858
CPCC12Q1/6858C12Q2535/137C12Q2545/101C12Q2545/113C12Q2531/113C12Q2561/101
Inventor 陆军
Owner 上海佰臻生物科技有限公司
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