Detection kit for human survival motor neuron genes SMN1 and SMN2, and detection method

Abstract

The invention discloses a detection kit for human survival motor neuron genes SMN1 and SMN2, and a method. The kit comprises an SMN1 main reaction liquid, an SMN2 main reaction liquid, an enzyme mixed liquid, 0-copy quality control, SMN1 single-copy quality control, SMN1 two-copy quality control, SMN2 single-copy quality control and SMN2 two-copy quality control, wherein the SMN1 main reaction liquid comprises a detection primer of the gene SMN1 and an inner reference primer of the SMN1; and the SMN2 main reaction liquid comprises a detection primer of the gene SMN2 and an inner reference primer of the SMN2. According to the kit, a PCR melting curve method is adopted, the change of a fluorescence signal value during a double-strand DNA melting process is detected in real time, target genes and inner reference genes generate different dissolution peaks according to the difference of different amplification product quantities, and then the copy numbers of the genes SMN1 and SMN2 are judged through software processing. The kit and the method which are disclosed by the invention are high in detection result accuracy, high in sensitivity, less in detection time consumption and low in cost.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a kit and a detection method for detecting the copy numbers of human motor neuron survival genes SMN1 and SMN2 by a PCR method. Background technique [0002] Spinal muscular atrophy (SMA) is one of the most common fatal neuromuscular diseases, which is caused by the degeneration of motor neurons in the anterior horn cells of the spinal cord. Hereditary neuromuscular disorder characterized by weakness, paralysis, and atrophy. The age of onset of SMA can range from before birth to adolescence. The main clinical symptoms and signs include symmetrical and progressive muscle strength, decreased muscle tone, and weakened or absent tendon reflexes involving the proximal muscles of the extremities. Atrophy and weakness, walking duck steps appear. In severe cases, the proximal muscles of the upper limbs are weak, and it is difficult to lift the shoulders. Intellect and sensations were ...

AI Technical Summary

Problems solved by technology

However, fluorescent dyes such as SYBR Green I have no sequence specificity, and cannot distinguish the difference in fluorescence signals generated by amplified products, primer dimers and non-specific products.

Whether the difference in amplification efficiency between the target gene and the internal reference gene is stable and consistent with the mathematical basis of the 2-△△Ct relative quantitative algorithm is still to be verified, so the reliability of this method in carrier detection is still open to question

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