Primer, probe and kit for detecting gene locus mutation, and using methods

Abstract

The invention discloses a primer, a probe and a kit for detecting gene locus mutation, and using methods. Primer sequences are shown as SEQ ID NO:1 and SEQ ID NO:2. Probe sequences are shown as SEQ ID NO:3 and SEQ ID NO:4. The kit comprises the primer, the probe, real-time fluorescence quantification PCR Mix reagents, deionized water, bottles or tubes which are used for separating the reagents, and a packaging box which is used for centralized packaging. According to the primer, the probe and the kit, the JAK2 gene V617F locus mutation can be quickly detected, the detection sensitivity is high, the probability of false positive or false negative is small, the detection steps are simple, the time consumption is low, and required clinical data can be obtained immediately.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to gene mutation detection primers, probes and a kit, in particular to a primer, a probe, a kit for detecting the V617F site mutation of the JAK2 gene and a use method thereof. Background technique [0002] The JAK2 gene belongs to the JAK gene family and encodes a JAK tyrosine protein kinase. Some growth factors and most cytokines can activate signal transducers and activators of transcription (STAT) through JAK, thereby affecting the transcriptional regulation of genes. The JAK-STAT signal transduction pathway is involved in many important biological processes such as cell proliferation, differentiation, apoptosis and immune regulation. JAK2 can mediate the signal transduction of various cytokines and growth factors, including erythropoietin (EPO), interleukin (IL-3), growth factor (GH), thrombopoietin (TPO), granulocyte-macrophage Colony-stimulating factor (GM-CSF) and g...

AI Technical Summary

Problems solved by technology

The conventional DNA sequencing method is to extract DNA from whole blood granulocytes, and directly sequence it after PCR amplification. However, because the mutation is acquired and limited to the myeloid lineage, the sensitivity of this detection method is only 20-30%. %, the detection sensitivity is low, and the test procedure is cumbersome, time-consuming, and costly; the ordinary qualitative PCR method is to design special primers, amplify the internal reference and the mutant fragment at the same time, and then observe the detection results by ordinary electrophoresis running gel, the sensitivity of this method The sensitivity is 20-30%, the detection sensitivity is low, and the detection results need to be read through the electrophoresis, which is highly subjective and prone to false positives or false negatives

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