Kit and primer pair group for detecting drug-induced deafness susceptibility gene mutation site

Abstract

The invention relates to the field of gene detection, in particular to a kit and a primer pair group for detecting drug-induced deafness susceptibility gene mutation sites, wherein the kit and the primer pair group can be used for detecting aminoglycoside drug susceptibility genes in mitochondria. A specific primer amplification and real-time fluorescence PCR technology is adopted, a new primer pair group is designed aiming at mutant nucleic acid to be detected, the primer is extended to product a specific PCR product when the mutant nucleic acid exists, a fluorescent substance is added into each reaction system, the fluorescent substance is bonded to the PCR product after amplification is completed, the produced melting peak is analyzed by adopting a melting curve, whether a corresponding mutant type exists on the nucleic acid or not is judged through the existence of the specific melting peak, and the specificity is high; the detection of two mutation sites can be completed by using a single tube and a single channel only, the requirements on instruments are low and the operation is simple to perform.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a kit and a special primer pair for detecting drug-induced deafness susceptibility gene mutation sites by using fluorescent PCR. Background technique [0002] Aminoglycoside antibiotics (streptomycin, gentamicin, kanamycin, sisomicin, tobramycin, etc.) cause protein translation errors by directly binding to 16S rRNA of the 30S subunit in bacterial ribosomes Or the premature termination of protein synthesis produces antibacterial effect. And because of its high-efficiency and broad-spectrum antibacterial effect, it is widely used clinically for the treatment of Gram-negative and positive bacterial infections, but this type of drug has serious ototoxic side effects, and can cause irreversible damage to hearing when used, and even " A needle for deafness". [0003] In recent years, it has been found that this susceptibility is usually maternally inherited and is associated with the A...

AI Technical Summary

Problems solved by technology

[0004] At present, the detection techniques for the above-mentioned two site mutations related to drug-induced deafness mainly include direct sequencing method, gene chip method, enzyme digestion method, and fluorescent probe method. High, complex interpretation or easy to cause cross-contamination, false positives and low sensitivity, only a single site detection and other shortcomings

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