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A kind of back-shaped lna probe and application thereof for point mutation detection

A technology of probes and probe sets, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbiological determination/inspection, etc., can solve the problems of insufficient detection specificity, low sensitivity of mutation detection, difficulty in meeting clinical needs, etc. To achieve simple and effective design, improve treatment effect, and improve specificity

Active Publication Date: 2021-08-24
嘉兴雅康博生物技术有限公司 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a back-shaped LNA probe for point mutation detection and its application, which solves the problem of insufficient detection specificity and low mutation detection sensitivity, and it is difficult to meet the current clinical needs

Method used

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  • A kind of back-shaped lna probe and application thereof for point mutation detection
  • A kind of back-shaped lna probe and application thereof for point mutation detection
  • A kind of back-shaped lna probe and application thereof for point mutation detection

Examples

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Effect test

Embodiment 1

[0027] Example 1: The detection of human CYP2C9 gene polymorphism using the back-shaped LNA probe system

[0028] The main enzymes that metabolize drugs in the human body are cytochrome P450 proteins (CYP), which are a class of monooxygenases that mainly exist in the liver and intestines. Metabolism of source substances (including most clinical drugs). P450 enzymes can transfer electrons to oxidize heterologous substances through iron ions in non-covalently bonded heme, enhance the water solubility of heterologous substances, and make them easier to excrete.

[0029] There are multiple subfamilies of CYP, among which CYP2C9 (Cytochrome P450 2C9) is an important member of the second subfamily, accounting for 20% of the total P450 protein in liver microsomal. CYP2C9 can hydroxylate and metabolize many drugs with different properties, mainly acidic substrates. According to statistics, about 16% of clinical drugs are metabolized by CYP2C9.

[0030] There are many mutant types o...

Embodiment 2

[0046]Example 2: The detection of the T790M mutation of the human EGFR gene by the back-shaped LNA probe system

[0047] Epidermal growth factor receptor (EGFR) is located on human chromosome 7, consisting of 30 exons, and is the expression product of the proto-oncogene ERBB1. EGFR mainly transmits signals to the nucleus through two pathways, one is Ras→Raf→MEK→ERK pathway; the other is PI3K→AKT→mTOR pathway. When the signal is transmitted to the nucleus, it will cause an increase in the transcription level of genes in the nucleus, resulting in cell proliferation and transformation. Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) are small molecule inhibitors targeting EGFR targets. Multiple clinical trials have confirmed that advanced NSCLC patients with EGFR mutations can benefit significantly from EGFR-TKIs. However, most patients will develop drug resistance after taking EGFR-TKIs for a period of time, and about 50% of drug-resistant patients can d...

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Abstract

The invention belongs to the field of biotechnology, and provides a back-shaped LNA probe for point mutation detection and applications thereof. The probe is an oligonucleotide with a length of 25 to 30 bp; the 5' end of the oligonucleotide is Label the luminescent group, label the quencher group at the 3' end, and make an LNA label on a base in the middle. The invention is suitable for single-base mutation detection, with simple and effective design, and is suitable for mainstream detection platforms on the market; the specificity of mutation detection is improved, the detection result of drug gene polymorphism is more accurate, and the effect of drug treatment is improved; the sensitivity of mutation detection is improved, Provide solutions for companion diagnosis and efficacy testing of tumor drugs.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an epsilon LNA probe for detecting point mutations and an application thereof. Background technique [0002] Polymerase chain reaction (PCR) technology is a simple, rapid, highly sensitive and specific gene amplification method, and its process usually includes 20-50 cycles consisting of three steps of denaturation, annealing and extension. It can specifically amplify the target nucleic acid sequence to several million times within 1.5-3 hours. In recent years, due to the appearance of real-time PCR instrument, gel electrophoresis analysis is no longer required for ordinary PCR, so post-PCR operations are not necessary, and PCR pollution is eliminated. Therefore, real-time PCR technology is more and more widely used in clinical practice. The most widely used in real-time fluorescent PCR detection technology is labeled fluorescent nucleic acid probes, such as Taqman prob...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q2525/301C12Q2531/113C12Q2563/107
Inventor 童超陈智林魏莎莎高幼冷
Owner 嘉兴雅康博生物技术有限公司
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