Detection kit and detection method for APL drug resistance gene mutation

Abstract

The embodiment of the invention provides a detection kit for APL drug resistance gene mutation and a detection method using the kit. The kit contains specific primers designed respectively for a PML / RARalpha fusion gene and protein mutational hotspot regains of PML and RARalpha, the primers are used for amplification of the PML / RARalpha fusion gene through PCR first and then respectively for amplification of the mutational hotspot regains of the PML and RARalpha, and finally sequencing analysis is carried out on target fragments in the APL sample amplification product to obtain the gene mutation condition of the APL sample. The kit and detection method provided by the invention can realize detection of mutation of three kinds of isomer PML / RARalpha fusion genes once, prevent S type and V type isomers from being undetected, and save the reagent cost at the same time; and by analyzing the gene mutation detection result acquired by the detection method provided by the embodiment of the invention, the suppression resistance caused by PML / RARalpha gene mutation to medicines such as As2O3 can be clinically revealed.

Description

technical field [0001] The invention belongs to the technical field of medicine and molecular biology, in particular to a detection kit for drug resistance gene mutation of acute promyelocytic leukemia (APL), which can be used to detect PML in PML / RARα fusion gene and mutations in the RARα region. The present invention also provides a method for detecting APL drug resistance gene mutation using the kit. Background technique [0002] APL is a special subtype of acute myelocytic leukemia (AML), accounting for about 10% to 15% of the total number of AML patients, and its morphological classification is M3 type. More than 98% of APL patients have a characteristic chromosomal translocation t(15;17)(q22;q12-21), which results in the combination of the promyelocytic leukemia (PML) gene on chromosome 15 and the dimension on chromosome 17. Fusion of the formate receptor alpha (RARα) gene to form a new PML / RARα fusion gene results in an arrest of granulocyte development at the promy...

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Problems solved by technology

[0003] The current international and domestic guidelines for the first-line treatment of APL are the use of arsenic trioxide (As 2 o 3 ) and all-trans retinoic acid (Tretinoin, Vitamin A Acid) or combination therapy has achieved great results, and the prognosis of APL patients has been significantly improved, and the average survival time of patients has been extended from 1 to 2 years in the past to 5 to 7 years. However, APL is still an incurable tumor, because some patients relapse after a period of medication, and relapse again after combination therapy, and the final prognosis is poor

With the progress of the research, it was found that the reason for the relapse and final poor prognosis of APL patients after treatment was the mutation of the PML and RARα protein regions in the PML / RARα fusion gene, resulting in the expression of As 2 o 3 and retinoic acid drug resistance; due to the existence of three different PML / RARα isomers, the usual gene mutation detection cannot cover all isomers, and in vitro amplification of large fusion fragments is required in the detection, how to target The targeted design of fusion large fragment amplification primers and the comprehensive and efficient detection of mutations in the three fusion types have become difficult problems in existing technologies.

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