A kind of spinal muscular atrophy pathogenic gene detection kit and its application

Abstract

The invention discloses a biosensor-based detection kit for spinal muscular atrophy (SMA) pathogenic genes (SMN1 and SMN2). The kit selects SMA pathogenic genes SMN1 and SMN2 as detection modules, and is based on strand displacement reaction. and DNA conformational transition, combined with the specific recognition of the fluorescent dye NMM (N-methylporphyrin dipropionate IX) and G-quadruplex, to construct several fluorescent biosensors, which can specifically recognize the target molecules SMN1 and SMN2, realize Fluorescence and visualization distinguish single-base differences between SMN1 and SMN2. In addition, the method can be applied to the genetic diagnosis of SMA, thereby bringing help and breakthroughs to the clinical diagnosis and prenatal genetic diagnosis of SMA. Compared with the prior art, the detection method of the present invention is a label-free fluorescence detection method and an ultraviolet-visible light transmission irradiation method. In the application, it can be identified by the fluorescence pattern and color change.

Description

A kind of spinal muscular atrophy pathogenic gene detection kit and its application technical field The present invention relates to DNA molecule detection and application, be specifically related to a kind of fluorescence biosensing based on DNA conformational transition The construction of the device and its application in the gene detection of spinal muscular atrophy pathogenic genes SMN1 and SMN2. Background technique Spinal muscular atrophy (Spinal Muscular Atrophy, SMA) is a kind of autosomal recessive genetic disease, with spinal cord Degeneration of motor neurons in the anterior horn of the spinal cord is a pathological feature. The incidence rate is about 1:6000~1:10000, and the incidence of carriers About 1:40, it is one of the main causes of death in children, and there is no effective treatment method. Given the severity of SMA, its Great grief for the patient's family. The main pathogenic gene of SMA is motor neuron survival gene (survival motor neur...

AI Technical Summary

Problems solved by technology

[0005] At present, there are many methods for the diagnosis of SMA, but most of them rely on gel electrophoresis to analyze DNA conformation or DNA sequencing, which has disadvantages such as poor specificity, low sensitivity, complicated operation, and high cost of detection.

For example, the most widely used restriction fragment length polymorphism analysis (PCR-restriction fragment length Polymorphism, PCR-RFLP) cannot detect the heterozygous deletion of SMN1, nor can it detect the copy number of SMN1, so it cannot distinguish between normal people and carriers Those who; Denaturing High-performance Liquid ChromatographHy (PCR-DHPLC), through the elution peak to determine whether there are different copies of base differences, still need to combine DNA sequencing to determine whether there is a deletion of SMN1; not yet Commercially applicable software tools have been developed for objective calculations, and peak shape diagnosis is difficult

Fluorescent real-time quantitative PCR technology can detect the copy number of SMN1, but the procedure of this method is complex, the required fluorescent-labeled probes are relatively expensive, the detection cost is high, and the result analysis is difficult.

Images