Primers, probes and kits for quantitative detection of spinal muscular atrophy genes

Abstract

The invention relates to the field of gene detection, and particularly relates to a primer, a probe and a kit for fluorescent quantitative detection on genes of spinal muscular atrophy (SMA). The kit comprises the primer and the probe for fluorescent quantitative detection on genes of SMA, wherein the primer includes 9 pairs of primers; the probe includes 15 probes. According to the primer, the probe and the kit for fluorescent quantitative detection on genes of SMA, a fluorescent quantitative PCR technology is utilized, common mutation site detection which is not found in existing patent technologies is added, so that, 16 types of SMA patients caused by SMN gene deletion, transformation and site mutation can be specifically detected in one time, and classification of SMA patients can be realized.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a primer, a probe and a kit for fluorescent quantitative detection of spinal muscular atrophy genes. Background technique [0002] Spinal muscular atrophy (SMA) is a relatively common genetic disease, and SMA with onset in children is autosomal recessive. The incidence rate of carriers is 1 / 35-1 / 80. It was recently reported that the carrier frequency of the SMA pathogenic gene in the Chinese population is 1 / 42. Chromosomal genetic disease is second only to cystic fibrosis. The Spinal Muscular Atrophy International Association is divided into 4 types according to the age of onset and clinical manifestations of SMA patients: type I (severe type), type II (intermediate type), type III (mild type) and type IV (adult type). [0003] At present, the methods commonly used clinically to assist the detection of spinal muscular atrophy mainly include electromyography, histochemical staining...

AI Technical Summary

Problems solved by technology

[0007] The patent "Spinal Muscular Atrophy-Related Gene Mutation Detection Method, Related Detection Probe Composition and Detection Kit, and Related Applications" only detects exon 7 and exon 8 of SMN1, and does not have the penetrance of SMN2 The detection of exon 7 means that only patients with SMA can be identified, but the severity of SMA patients is not identified, which has no guiding significance for the quality of life of patients, and the detection of exon 7 and exon 8 is divided , which indirectly increases the workload and detection cost; the patent "A Fluorescent Quantitative PCR Kit for Diagnosing Human Spinal Muscular Atrophy" detects SMN1 and SMN2, and also adds the NAIP gene for detection, although it increases the SMA disease Genotyping for detection, but this method uses Taqman to detect only a few single-base differences in SMN1 and SMN2, which is likely to cause non-specificity, and has a very high risk in clinical detection; in summary, the above methods can only be realized Detection of patients with deletional SMA; there are still patients with non-deletion SMA who are not covered by the protocol

[0008] In the prior art, there are few detection sites for spinal muscular atrophy pathogenic genes, and it is impossible to classify patients with SMAI type (severe type), type II (intermediate type), and type III (light type), and most of the fluorescence quantitative Using single internal reference standardization, there are certain subjective factors in the judgment of quantitative test results, which leads to the results not accurately reflecting the real situation of the sample, and increases the risk of clinical testing, especially prenatal diagnosis; while using MLPA technology takes a long time to operate and requires multiple steps to open the cover The operation increases the risk of contamination, the detection cost is high, and the detection of mutation points cannot be realized, so it cannot meet the needs of rapid clinical diagnosis of SMA and a wide detection range.

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