Kit for detecting K-ras gene mutation and detection method with kit

Abstract

The invention discloses a kit for detecting K-ras gene mutation and a detection method with the kit. The kit comprises an amplification primer pair, a TaqMan-MGB (minor groove binder) probe and PNA (pentose nucleic acid), wherein the sequences of amplification primers are shown as SEQ ID NO:1 and SEQ ID NO:2; and the sequence of the TaqMan-MGB probe is shown as SEQ ID NO:3; and the sequence of PNA is shown as SEQ ID NO:4. The detection method comprises the following steps of: extracting DNA (deoxyribonucleic acid) of a sample to be detected; preparing K-ras wild and mutation plasmid standard products; and detecting the K-ras gene mutation, performing qualitative judgment on the K-ras gene mutation, drawing a standard curve, performing quantitative analysis on the mutation, and the like. The kit has the characteristics of quickness, simplicity, convenience, high sensitivity and high flux, and can be used for performing qualitative or quantitative detection on the K-ras gene mutation of clinical samples of tissue, urea, serum or plasma.

Description

technical field [0001] The invention belongs to the technical field of molecular detection, and in particular relates to a kit for quantitatively detecting whether K-ras mutant gene is contained in tissue, urine, serum or plasma and a detection method thereof. Background technique [0002] The mutational activation of K-ras is one of the main reasons for the malignant transformation of various human tumor cells. It is like a molecular switch, which can control and regulate the path of cell growth in normal conditions; when it is abnormal, it will lead to uncontrolled cell proliferation and prevent cell self-destruction. K-ras gene mutations mostly occur at the 12th and 13th codons, and this abnormality is found in pancreatic cancer (75%-95%), colorectal cancer (40%-50%) and lung cancer (20%-30%) and other tumors The incidence rate in tissues is relatively high, and K-ras gene mutations can often be detected in the serum or plasma of patients with these tumors. [0003] At p...

AI Technical Summary

Problems solved by technology

At present, detection methods with a sensitivity of >5% such as sequencing are still mostly used clinically, and the detection sensitivity of the mutant gene-specific amplification PCR method (ARMS) is generally between 1% and 5%, which is not well applicable to serum or plasma. K-ras gene mutation detection

In the June 2012 issue of Nature, two research teams from Italy and the United States reported that a considerable part of colorectal cancer’s drug resistance to targeted drugs is related to the presence of K-ras gene mutations in the patient’s tumor tissue. Because the mutation rate of K-ras gene in the tumor tissue of these patients is low, it is difficult to detect it by conventional methods such as sequencing.

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