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KRAS gene mutation multiple detection primer probe and kit thereof

A multiplex detection, primer-probe technology, applied in the direction of DNA/RNA fragment, recombinant DNA technology, microbial determination/inspection, etc., can solve the problem of limiting ctDNA detection, and achieve the effect of accurate results

Pending Publication Date: 2022-03-18
普瑞斯新(上海)生物医疗科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moderate sensitivity of mutation detection methods currently used in clinical practice limits detection of ctDNA
Sensitivity thresholds for traditional methods are around 1%, but ctDNA may represent only a small fraction of total circulating DNA

Method used

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  • KRAS gene mutation multiple detection primer probe and kit thereof
  • KRAS gene mutation multiple detection primer probe and kit thereof
  • KRAS gene mutation multiple detection primer probe and kit thereof

Examples

Experimental program
Comparison scheme
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Embodiment

[0030] The KRAS gene mutation multiple detection kit of this embodiment includes upstream primer F-E2, downstream primer R-E2, G12 site wild-type probe WP-G12, G13 site wild-type probe WP-G13, to detect KRAS gene 2 Mutant probe MP-G12D, mutant probe MP-G12C, mutant probe MP-G12V, mutant probe MP-G12S, mutant probe MP-G12A, mutant Probe MP-G12R, a mutant probe MP-G13D for detecting mutations at the G13 site of exon 2 of the KRAS gene, a plasmid with mutations in the KRAS genes G12D, G12C, G12V, G12S, G12A, G12R and G13D.

[0031] The primers, probes and plasmids involved in this example were all designed by the laboratory technicians of Price New (Shanghai) Biomedical Technology Co., Ltd., and the synthesis unit was completed by Nanjing GenScript Biotechnology Co., Ltd. The primers and The nucleotide sequences of the probes are shown in Table 1.

[0032] Table 1 Primer probe characteristic table

[0033]

[0034] The primers are conventional primers, the probes are modifie...

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Abstract

The invention relates to a KRAS gene mutation multiple detection primer probe and a kit thereof. The KRAS gene mutation multiple detection primer probe comprises an upstream primer, a downstream primer, a G12 site wild type probe WP-G12, a G13 site wild type probe WP-G13, a mutant probe MP-G12D, a mutant probe MP-G12C, a mutant probe MP-G12V, a mutant probe MP-G12S, a mutant probe MP-G12A and a mutant probe MP-G12R, wherein the mutant probe MP-G12D, the mutant probe MP-G12C, the mutant probe MP-G12V, the mutant probe MP-G12S, the mutant probe MP-G12A and the mutant probe MP-G12R are used for detecting mutation of the G12 site of a second exon of a KRAS gene; and the mutant probe MP-G13D is used for detecting the mutation of the G13 site of the second exon of the KRAS gene. The KRAS gene mutation multiple detection primer probe and the kit thereof have the advantages of rapidness, high efficiency, simple preparation of a standard substance, high specificity and sensitivity of a detection system, high utilization rate of free DNA, and high accuracy.

Description

technical field [0001] The invention relates to a KRAS gene mutation detection product, which belongs to the field of biotechnology. Background technique [0002] Among all NSCLC treatment-prognostic biomarkers, KRAS mutations are characterized by their high frequency of events (approximately 30% of adenocarcinomas), KRAS negative predicts a better prognosis, and patients with positive mutations lack effective targeted therapy. Mutations in the KRAS gene are present in 40% of colorectal adenocarcinomas and are predictive markers of unresponsiveness to anti-epidermal growth factor receptor (anti-EGFR) 9 antibodies. The National Comprehensive Cancer Network (NCCN) of the United States clearly states that all patients with metastatic colorectal cancer should be tested for KRAS gene status, and it is not recommended to use Tarceva for molecular targeted therapy in non-small cell lung cancer patients with KRAS gene mutations. Recent preclinical findings and early trials suggest ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/156C12Q2600/16C12Q2600/166Y02A50/30
Inventor 洪专潘文健
Owner 普瑞斯新(上海)生物医疗科技有限公司
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