Spinal muscular atrophy pathogenic gene detection kit based on melting curve analysis

Abstract

The invention relates to a spinal muscular atrophy pathogenic gene detection kit based ona melting curve analysis, and relates to a pathogenic gene detection kit, which comprises an upstream primer F1 and a downstream primer R1 for amplifyingexon7 of SMN1 and SMN2 genes; an upstream primer F2 and a downstream primer R2 for amplifying exon4 of an internal reference CFTR gene; and a fluorescent probe for detection. In a single-tube PCR system, the copy number of exon7 of the SMN1 gene as a main pathogenic gene of SMA can be quantitatively detected, the sample genotype can be known by a fluorescence PCR melting curve analysis after PCR amplification is finished, the whole operation is completed in 2 to 3h, is simple and rapid, and is short in time-consuming; the homogeneous detection and closed tube operation are carried out; the detection flux is high; the detection specificity is high, and the results are easy to interpret. The detection kit can be rapidly and convenientlyapplied to large-scale population screening of the SMA pathogenicgene, and is especially suitable for prenatal, premaritaland pre-pregnancy screening and genetic diagnosis of patients.

Description

technical field [0001] The invention relates to a pathogenic gene detection kit, in particular to a spinal muscular atrophy pathogenic gene detection kit based on melting curve analysis. Background technique [0002] Spinal muscular atrophy (SMA) refers to a group of diseases with progressive skeletal muscle weakness and atrophy caused by the degeneration of the anterior horn cells of the spinal cord. It is an autosomal recessive genetic disease with an incidence rate of 1 / 6000~1 / 10000. The clinical manifestations are progressive, symmetrical muscle atrophy and muscle weakness. So far, there is no effective treatment for SMA, and the prognosis is mainly related to the type of disease. Type I patients develop symptoms 6 months after birth, and generally survive within 2 years; Within 5 years of age, and type III patients can survive to adults, their disease progresses slowly, and eventually die of respiratory muscle paralysis or systemic failure. [0003] The currently di...

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Problems solved by technology

[0005] The SMA gene diagnosis methods adopted in the prior art mainly include polymerase chain reaction-restriction fragment length polymorphism analysis technology, allele-specific amplification, multiplex ligation probe-dependent amplification technology (MLPA) etc., However, restriction fragment length polymorphism analysis and allele-specific amplification can only qualitatively detect whether a patient has SMN1 gene homozygous deletion, and cannot distinguish SMN1 pathogenic gene carriers. Although MLPA can carry out carrier screening It is not suitable for large-scale clinical screening because of its high technical cost, high equipment requirements, complicated operation and long time-consuming

[0006] Although patents such as CN 105039318A, CN 103789440A, and CN 104480206A have also proposed the use of fluorescent PCR and other technologies to detect spinal muscular atrophy-related gene mutations, but these technical solutions generally have low accuracy, poor reliability, and high cost. Weaknesses in prenatal diagnosis and mass population screening

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