Primer probe for detecting mutation of KRAS gene and kit thereof

A technology for detecting kits and detection primers, applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial measurement/inspection, etc., can solve problems such as difficult, limited length, difficult simultaneous detection of multiple sites, etc. To achieve the effect of reducing costs, avoiding suppression, and avoiding adverse effects

Inactive Publication Date: 2017-08-08
上海赛安生物医药科技股份有限公司
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

Due to the limitation of hardware equipment, it is difficult to realize simultaneous detection of multiple sites, and the dPCR multiplex

Method used

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  • Primer probe for detecting mutation of KRAS gene and kit thereof
  • Primer probe for detecting mutation of KRAS gene and kit thereof
  • Primer probe for detecting mutation of KRAS gene and kit thereof

Examples

Experimental program
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Embodiment 1

[0031] 1. The composition of the kit.

[0032] The KRAS gene mutation detection kit of this embodiment includes a digital PCR premix, a droplet stabilizer, a primer-probe mixture, a histone deacetylase solution and a positive control. The components of the kit are shown in Table 1.

[0033] Table 1 Kit composition list

[0034] Element pack Reagent company Digital PCR Master Mix 1 tube Life technologies droplet stabilizer 1 tube Raindance technologies Primer Probe Mix 1 tube Bailige positive control 1 tube —— Histone deacetylase 1 tube Beijing Solaibao Technology Co., Ltd.

[0035] The description of the kit components in the above Table 1 is as follows:

[0036] 1) The main components of the digital PCR master mix include DNA polymerase, ultrapure water, dNTPs mixture, reference fluorescent ROX, buffer solution, etc. (provided by Life technologies, product number: 1508099).

[0037] 2) The main component of the d...

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Abstract

The invention relates to a primer probe for detecting mutation of a KRAS gene and a kit thereof. The kit comprises digital PCR (polymerase chain reaction) premix liquid, a liquid droplet stabilizer and primer probe mixing liquid, wherein the digital PCR premix liquid is used for preparing digital PCR reaction liquid; the primer probe mixing liquid comprises upstream and downstream primers A for detecting a 12th codon, a mutation probe A for a G12R site of the 12th codon, a mutation probe B for a G12V site of the 12th codon, a mutation probe C for a G12D site of the 12th codon, a mutation probe D for a G12A site of the 12th codon, a mutation probe E for a G12S site of the 12th codon, and a mutation probe F for a G12C site of the 12th codon; upstream and downstream primers B for detecting a 13th codon, and a mutation probe G for a G13D site of the 13th codon; upstream and downstream primers C for detecting a 61st codon, and a mutation probe H for a Q61L site of the 61st codon. The kit for detecting the mutation of the KRAS gene has the advantages that the sensitivity and specificity are high, the sample demand amount is small, and multiplex PCR detection can be rapidly and accurately performed.

Description

technical field [0001] The invention relates to a primer, a probe and a detection product for detecting KRAS gene mutation, belonging to the field of biotechnology. Background technique [0002] RAS gene mutations are considered to be closely related to human tumorigenesis. Mutations of codons 12 and 13 of the KRAS gene in the RAS family have a high detection rate in colorectal cancer and pancreatic cancer, and are considered to be directly related to tumor occurrence, recurrence and patient prognosis. Carrying the KRAS mutation gene, its changes at the gene level with CDKN2A, SMAD4, and TP53 are considered to be directly related to the occurrence and development of pancreatic cancer. Monitoring the changes in KRAS mutations in vivo helps to understand the relationship between gene changes and disease progression . [0003] Exploring the mechanism of KRAS drug resistance and predicting the prognosis of patients has become a feasible way. Now it is clear that tumor cells wi...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/106C12Q2600/118C12Q2600/156
Inventor 赵新泰王明赵会
Owner 上海赛安生物医药科技股份有限公司
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