A kind of smn1 gene mutation detection kit and its application

Abstract

The invention relates to the field of molecular biology diagnosis, in particular to a detection kit for SMN1 gene mutation. In a tube of long-fragment PCR reaction, three kinds of primers are used to specifically amplify the gene sequence of the SMN1 gene protein coding region, and the SMN gene sequencing primer set is used for direct sequencing. Only one PCR experiment can detect the SMN1 gene mutation, including : Point mutations, deletion mutations and insertion mutations, etc., the detection technology process is simple and easy to achieve standardization.

Description

technical field [0001] The invention relates to the field of molecular biology diagnosis, in particular to a detection kit for SMN1 gene mutation, and the mutation types mainly include: point mutation, deletion mutation and insertion mutation. Background technique [0002] Spinal muscular atrophy (SMA) is a disease in which the degeneration of motor neurons in the anterior horn of the spinal cord leads to muscle weakness, muscle atrophy and paralysis. ), Type III (mild) and Type IV (adult). Clinical diagnosis is made according to the age of onset, clinical manifestations, and degree of disease progression. At the same time, relevant auxiliary examination methods include electromyography, muscle enzymes, and muscle biopsy, combined with family history and SMN1 gene analysis to confirm the diagnosis. SMA type Ⅰ is the most severe subtype, with onset within 6 months of birth, and the general survival period is within 2 years; SMA type Ⅱ is the moderate type, with onset in pati...

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Problems solved by technology

At present, there are several methods: (1) RT-PCR combined with cloning and sequencing, but the cloning technology is cumbersome and time-consuming, and it is not easy to realize the standardized detection process for the detection of compound heterozygous mutations in the SMN1 gene (2) Extract RNA and reuse SMN1-specific primers are reverse-transcribed into cDNA, followed by PCR and sequencing, which is also cumbersome and time-consuming; (3) Another common method is to carry out SMN1 gene specificity through LR-PCR (Long Range PCR). Amplify, and then carry out the second PCR amplification and sequencing for each exon to confirm the position of the SMN1 gene point mutation, but because the SMN1 and SMN2 genes have only 5 nucleotide differences, the SMN1 gene is specifically amplified It is difficult, but interfering with the detection and interpretation of compound heterozygous mutations in the SMN1 gene

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