Reagent and kit for detecting CALR gene mutation

Abstract

The invention discloses a reagent and a kit for detecting CALR gene mutation. The reagent comprises a CALR gene specific primer pair, a probe and a CLAMP nucleic acid sequence, the specific primer pair can specifically amplify the CALR gene containing the mutation region; the CLAMP nucleic acid sequence is a 3'-terminal phosphorylated single strand which is only completely complementary with the CALR wild type gene, the CLAMP nucleic acid sequence is complementarily combined with one DNA strand of the CALR gene, and the complementary combination region covers the 3'-terminal of one primer in the specific primer pair and covers the mutation region; the CLAMP nucleic acid sequence, the specific probe and the primer covered by the CLAMP nucleic acid sequence are combined on the same DNA chainof the CALR gene. According to the reagent disclosed by the invention, wild type gene amplification is inhibited through competition between the primers and the CLAMP nucleic acid sequence, so that the sensitivity of CALR gene mutation detection is improved, the false negative rate is reduced, and the detection rate is increased.

Description

technical field [0001] The application relates to the field of gene mutation detection, in particular to a reagent and kit for detecting CALR gene mutation. Background technique [0002] Myeloproliferative neoplasms (myeloproliferative meoplasm, MPN) is a group of malignant tumors originating from pluripotent hematopoietic stem cells, characterized by abnormal clonal proliferation of one or more lines of bone marrow hematopoietic cells, including polycythemia vera (polycythemia, PV), primary Essential thrombocythemia (essential thrombocthemia, ET), primary myelofibrosis (primary myelofibrosis, PMF). [0003] Studies have shown that 97% of PV, 50%-60% of ET and PMF patients have JAK2 V617F gene mutation, MPL515 gene mutation can be found in 3%-5% of ET and PMF patients carrying wild-type JAK2 V617F gene, however, still About 35%-40% of ET and PMF patients have no abnormality of JAK2 V614F gene and MPL515 gene detected. In recent years, studies have pointed out that about 70...

AI Technical Summary

Problems solved by technology

However, for a large number of wild-type CALR genes mixed with a small amount of mutants, when there is cross-reaction between the wild-type gene and the mutant gene, the detection of the mutant CALR gene by ordinary fluorescent PCR method has low sensitivity, high false negative rate, and the problem of low detection rate

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