Primer and probe for screening spinal muscular atrophy (SMA) genes and using method of primer and probe

Abstract

The invention discloses a primer and a probe for screening spinal muscular atrophy (SMA) genes and a using method of the primer and probe, belonging to the technical field of biology. The invention discloses eight combinations of the primer and probe for screening survival motor neuron genes 1 (SMN1), and the primer and probe can be effectively applied to screening the SMN1. Meanwhile, the invention also discloses sixteen groups of combinations of the primer and probe for screening survival motor neuron genes 2 (SMN2), as well as application in fetal SMA gene screening by utilizing the primer and probe for screening the SMN1 and the primer and probe for screening the SMN2. According to the primer and probe provided by the invention, the SMA genotypes of adults and fetuses can be detected at high efficiency.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to primers, probes and application methods for human spinal muscular atrophy (SMA) gene screening. Background technique [0002] Spinal muscular atrophy (SMA) is a group of autosomal recessive genetic diseases characterized by degeneration of motor neurons in the anterior horn of the spinal cord. The clinical manifestations are progressive and symmetrical proximal limb and trunk muscles. Weakness, atrophy and paralysis, the incidence rate of newborns is 1 / 6000-1 / 10000, and the incidence rate of gene carriers in the normal population is 1 / 40-1 / 60, ranking second among fatal autosomal recessive genetic diseases , second only to cystic fibrosis. [0003] According to the age of onset and clinical manifestations, SMA can be divided into four types: (1) Type I SMA (acute type): Onset within 6 months after birth, generalized muscle weakness, hypotonia, children cannot sit and sta...

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Problems solved by technology

However, PCR-SSCP technology has high experimental requirements, and temperature changes during electrophoresis can easily cause false positive or false negative results by affecting the folding of single-stranded DNA, which limits its application in clinical laboratories; Due to the cumbersome operation steps of clinical gene detection, post-processing of PCR products is required, contamination is prone to occur during the operation, and there are false negative results caused by incomplete digestion and heteroduplexes during amplification that cannot be digested. It is also impossible to detect the loss of heterozygosity of the SMN1 gene, so it is impossible to distinguish between SMA pathogenic gene carriers and normal people; DHPLC method is used for SMA pathogenic gene detection, and can only detect heterozygous mutations. Due to the sensitivity of the detection system, it is easy to cause detection failure Or the repeatability of the results is not good, and because the copy number of the SMN gene fluctuates greatly, there are certain problems in data analysis; MLPA technology is applied to the detection of SMA pathogenic genes, which can not only detect the copy number of the SMN gene, but also determine the heterozygote and Homozygous deletion, in addition, due to its semi-quantitative effect, it can detect the carrier of the disease-causing gene, and has the advantages of high accuracy, good repeatability, simplicity and speed, etc. This method combined with DNA sequencing technology can make the result of genetic diagnosis more accurate Reliable, but MLPA also has its limitations: 1. It needs to accurately measure the concentration of DNA, and the sample is easily contaminated; 2. It cannot be used for the detection of single cells; 3. It cannot detect the balanced translocation of chromosomes

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