Primer composition, probe, kit and method for detecting JAK2 gene

Abstract

The invention provides a primer composition and probe for detecting a JAK2 gene, and is characterized in that the primer composition comprises a primer pair for detecting a mutant gene and a primer pair for detecting a wild type gene; the primer pair for detecting the mutant gene contains an upstream primer with a sequence shown as Seq ID No. 1 and a downstream primer with a sequence shown as SeqID No. 2; the primer pair for detecting the wild type gene comprises an upstream primer with a sequence shown as Seq ID No. 1 and a downstream primer with a sequence shown as Seq ID No. 3; the probe has a sequence as shown in Seq ID No.4; M at the 5' terminal of each primer is a nucleic acid sequence which is not complementary with the JAK2 gene and is used for increasing the GC content of each primer. According to the primer composition and the probe for detecting the JAK2 gene, the specific binding of the primer and the amplification template is enhanced, and when M is introduced into the amplification template, the Tm value of the complementary region of the primer and the JAK2 gene is increased, and effective annealing can be realized, so that the sensitivity of detecting JAK2 gene mutation is improved.

Description

technical field [0001] The present application relates to the field of gene mutation detection, in particular to a primer composition, probe, kit and method for detecting JAK2 gene. Background technique [0002] Myeloproliferative neoplasms (MPNs) are a group of malignant tumors originating from multipotent hematopoietic stem cells, characterized by abnormal clonal proliferation of one or more lines of bone marrow hematopoietic cells, including polycythemia vera (PV), primary Essential thrombocythemia (essential thrombocthemia, ET) and primary myelofibrosis (primary myelofibrosis, PMF), etc. [0003] Janus kinases (JAKs) are a family of non-receptor tyrosine protein kinases, currently including four members: JAK1, JAK2, JAK3, and TYK2. JAK kinases have highly homologous domains: at the carboxy-terminus there is an active kinase domain (JH1 domain of JAK homology) and a pseudokinase domain (JH2 domain) that catalyzes the "inactive state kinase" ). Studies have shown that t...

AI Technical Summary

Problems solved by technology

However, the sequencing method is expensive, complicated, and time-consuming, which is not conducive to rapid clinical diagnosis

The JAK2 gene mutation can also be detected by amplification-restricted mutation PCR (ARMS-PCR). Amplification-restricted mutation PCR can detect single-gene mutations under ideal PCR conditions, and can distinguish whether a JAK2V617F mutant individual is homozygous or heterozygous. There is a reliable method for mutations in MPNs patients, but the disadvantage of this method is that the sensitivity is low, only 1%-2%

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