Multiplex PCR primer and method for detecting gene fusion through combination of anchored nested multiplex PCR and high-throughput sequencing

Abstract

The invention provides a multiplex PCR primer detecting gene fusion through combination of anchored nested multiplex PCR and high-throughput sequencing. The multiplex PCR primer comprises a multiplex PCR specific primer group 1 and a multiplex PCR specific primer group 2, wherein the multiplex PCR specific primer group 1 consists of nucleotide sequences shown in SEQ ID NO:1-SEQ ID NO:32, and the multiplex PCR specific primer group 2 consists of nucleotide sequences shown in SEQ ID NO:33-SEQ ID NO:64. The invention further provides a method for detecting gene fusion through combination of anchored nested multiplex PCR and high-throughput sequencing. The detection method is high in universality and can be used for detecting fusion genes accurately and sensitively under the condition that structures of fusion genes are partially known or unknown.

Description

technical field [0001] The present invention relates to the field of detection, more specifically to a multiplex PCR primer and method for anchoring nested multiplex PCR combined with high-throughput sequencing to detect gene fusion Background technique [0002] Studies have shown that in mammals such as humans, gene fusions are one of the causes of certain diseases such as cancer. [0003] Taking mixed lineage-leukemia gene (MLL for short) as an example, MLL fusion protein can cause abnormal epigenetic modification of downstream genes, leading to activation of downstream oncogenes, thereby inducing leukemia. [0004] The fusion protein produced by ectopic MLL acts as an oncogene. In addition to the most common MLL-MLLT3 (acutemyeloid leukemia (AML) acute myeloid leukemia) and MLL-MLLT2 (AF4), there are MLL fusions with the following partner genes Genes: MLLT1 (ENL), MLLT10 (AF10), MLLT4 (AF6), ELL, etc. [0005] Among many MLL fusion genes, for acute myeloid leukemia (AML...

AI Technical Summary

Problems solved by technology

Limitations of this method: 1) It is necessary to know the gene that is fused with the MLL gene; 2) It is necessary to know the way the two genes are fused; 3) It is impossible or difficult to detect the fusion of different genes with MLL at the same time

However, this method has limitations: 1) The gene fused with the MLL gene must be known; 2) The chromosome position of the gene fused with MLL must be at a relatively short distance from MLL; 3) The occurrence of different genes with MLL cannot be detected simultaneously. fusion of

Therefore, this method must design different probes for different genes fused with the MLL gene, and cannot detect unknown MLL fusion genes, which will affect the detection rate of the MLL gene

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