Detection method of electrochemical acute leukemia gene Pax-5a based on enzyme assisted cyclic signal amplification

Abstract

Provided is a detection method of an electrochemical acute leukemia gene Pax-5a based on enzyme assisted cyclic signal amplification; the efficiency advantage of transcription termination and translation is controlled based on a hairpin DNA probe (HP). The target gene Pax-5a opens HP, and the replication template function of the HP is started. Under the synergistic effect of DNA polymerase and restriction endonuclease, on one hand, auxiliary DNA promotes the target gene Pax-5a to cyclically open the HP; on the other end, a large number of G-quadruplex sequences are produced. The G-quadruplex sequences are captured by a sulfhydryl-modified probe on the surface of a gold electrode due to the complementary action of bases, then a G-quadruplex / hemin complex is formed in the presence of hematinchloride and potassium ions, and hydrogen peroxide can be catalyzed to reduce so as to generate electrochemical signals; the change of the current has linear correlation relationship with the concentration of the target gene in a certain range. Based on the mechanism of enzyme assisted cyclic signal amplification of the hairpin DNA probe and a one-key electrochemical detection platform, high-sensitivity and high-selectivity detection of the target gene can be achieved.

Description

technical field [0001] The invention relates to the field of electrochemical biological analysis, in particular to a detection method for acute leukemia gene Pax-5a. Background technique [0002] As one of the most common human malignancies, acute lymphoblastic leukemia (ALL) has the highest prevalence among children. About 3,000 to 4,000 people are diagnosed with ALL each year in the United States, two-thirds of whom are children between the ages of 2 and 5. Pax-5 is the only paired-box (PAX) family member found in the hematopoietic system, which can be divided into Pax-5a, Pax-5b, Pax-5c, Pax-5d and Pax-5e. Among them, Pax-5a is the most important, and Pax-5 usually refers to Pax-5a. Pax-5 is very important for the differentiation and development of B cells, and its abnormal expression can cause B lymphocytic leukemia. However, there are few studies on the detection of variant Pax5 gene and the expression of this gene in clinical acute lymphoblastic leukemia patients. ...

AI Technical Summary

Problems solved by technology

[0006]However, electrochemical biosensors generally have certain limitations in terms of signal generation

On the one hand, when constructing electrochemical biosensors, appropriate electroactive substances need to be labeled, which may limit the detection sensitivity; on the other hand, general electrochemical layer-by-layer assembly reactions will continuously introduce interference substances on the electrode surface; thirdly, Incorporating appropriate signal amplification techniques into the construction of electrochemical biosensors directly at the electrode interface is an efficiency issue

Images